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potential probiotic thermophiles isolated from mice after compost ingestion.the oral administration of a compost produced by the fermentation of marine animals with thermophiles confers health benefits for fish and pigs. this study aimed to isolate the beneficial bacteria from this compost that would modulate the physiological conditions of host animals.201323311468
jerusalem artichoke powder: a useful material in producing high-optical-purity l-lactate using an efficient sugar-utilizing thermophilic bacillus coagulans strain.jerusalem artichoke is a low-requirement crop, which does not interfere with food chain, and is a promising carbon source for industrial fermentation. microbial conversion of such a renewable raw material to useful products, such as lactic acid, is an important objective in industrial biotechnology. in this study, high-optical-purity l-lactate was efficiently produced from the hydrolysates of jerusalem artichoke powder by a thermophilic bacterium, bacillus coagulans xzl4. high l-lactate producti ...201323306126
functional analysis of the comk protein of bacillus coagulans.the genes for dna uptake and recombination in bacilli are commonly regulated by the transcriptional factor comk. we have identified a comk homologue in bacillus coagulans, an industrial relevant organism that is recalcitrant for transformation. introduction of b. coagulans comk gene under its own promoter region into bacillus subtilis comk strain results in low transcriptional induction of the late competence gene comga, but lacking bistable expression. the promoter regions of b. coagulans comk ...201323301076
inoculation of pichia kudriavzevii rb1 degrades the organic acids present in raw compost material and accelerates composting.in this study, the yeast strain pichia kudriavzevii rb1 was used as an inoculum to accelerate organic matter degradation of rabbit food with added organic acids, which was used as a model food waste for composting. the rb1 strain rapidly degraded the organic acids present in the raw compost material, leading to an increase in ph beyond the neutral level, within 2 days. both mesophilic and thermophilic bacteria proliferated faster in the compost with rb1 inoculation than in that without inoculati ...201323886646
genome sequences of two morphologically distinct and thermophilic bacillus coagulans strains, h-1 and xzl9.two thermophilic bacillus coagulans strains, h-1 and xzl9, both of which were isolated from soils, have different morphological properties. strain xzl9 but not h-1 is an efficient pentose-utilizing producer of important platform compounds, such as l-lactic acid and 2,3-butanediol. here we announce the 2.86- and 3.43-mb sequences of their genomes.201323682151
structure and dynamics of the bacterial communities in fermentation of the traditional chinese post-fermented pu-erh tea revealed by 16s rrna gene clone library.microbes are thought to have key roles in the development of the special properties of post-fermented pu-erh tea (pu-erh shucha), a well-known traditional chinese tea; however, little is known about the bacteria during the fermentation. in this work, the structure and dynamics of the bacterial community involved in the production of pu-erh shucha were investigated using 16s rrna gene clone libraries constructed from samples collected on days zero (ld-0), 5 (ld-5), 10 (ld-10), 15 (ld-15) and 20 ( ...201323591759
screening of bacillus coagulans strains in lignin supplemented minimal medium with high throughput turbidity measurements.the aim of this study was to extend the options for screening and characterization of microorganism through kinetic growth parameters. in order to obtain data, automated turbidimetric measurements were accomplished to observe the response of strains of bacillus coagulans. for the characterization, it was decided to examine the influence of varying concentrations of lignin with respect to bacterial growth. different mathematical models are used for comparison: logistic, gompertz, baranyi and rich ...201428626663
effect of bacterial cell-free supernatants on infectivity of norovirus surrogates.bacterial metabolic products were evaluated for inhibitory effects on viral propagation in cell culture. cell-free supernatants (cfs) were prepared from growth of enterococcus faecalis atcc 19433, pseudomonas fluorescens atcc 13525, escherichia coli 08, staphylococcus epidermidis atcc 12228, bacillus subtilis 168, bacillus coagulans 185a, b. coagulans 7050, clostridium sporogenes pa3679, and a commercial probiotic mixture of lactobacillus acidophilus, lactobacillus rhamnosus, bifidobacterium bif ...201424406013
bacillus coagulans tolerance to 1-ethyl-3-methylimidazolium-based ionic liquids in aqueous and solid-state thermophilic culture.the use of ionic liquids (ils) to disrupt the recalcitrant structure of lignocellulose and make polysaccharides accessible to hydrolytic enzymes is an emerging technology for biomass pretreatment in lignocellulosic biofuel production. despite efforts to reclaim and recycle il from pretreated biomass, residual il can be inhibitory to microorganisms used for downstream fermentation. as a result, pathways for il tolerance are needed to improve the activity of fermentative organisms in the presence ...201424376258
antimicrobial and antioxidative activities in the bark extracts of sonneratia caseolaris, a mangrove plant.the present study deals with the phytochemical contents, antimicrobial and antioxidative activities of bark tissue of sonneratia caseolaris, a mangrove plant from sundarban estuary, india. phytochemical analyses revealed the presence of high amounts of phenolics, flavonoids, tannins, alkaloids and saponins. antimicrobial efficacies of various extracts of s. caseolaris were assessed by disc diffusion method against two gram-positive (bacillus subtilis and bacillus coagulans), two gram-negative (e ...201426417316
efficient production of l-lactic acid using co-feeding strategy based on cane molasses/glucose carbon sources.l-lactic acid is an important platform chemical, which ought to be produced under cost control to meet its huge demand. cane molasses, a waste from sugar manufacturing processes, is hopeful to be utilized as a cheap carbon source for l-lactic acid fermentation. considering that cane molasses contains nutrients and hazardous substances, efficient production of l-lactic acid was developed by using a co-feeding strategy based on the utilization of cane molasses/glucose carbon sources. based on the ...201424333698
open fermentative production of l-lactic acid with high optical purity by thermophilic bacillus coagulans using excess sludge as nutrient.the development of a low-cost polymer-grade l-lactic acid production process was achieved in this study. excess sludge hydrolyzate (esh) was chosen as nutrient source for the objective of reducing nutrient cost in lactic acid production. 1% of esh had high performance in lactic acid production relative to 2g/l yeast extract (ye) while the production cost of esh was much lower than that of ye, indicating esh was a promising substitute of ye. by employing a thermophilic strain of bacillus coagulan ...201424201025
isolation of thermophilic l-lactic acid producing bacteria showing homo-fermentative manner under high aeration condition.by applying non-sterile open fermentation of food waste, various thermotolerant l-lactic acid-producing bacteria were isolated and identified. the predominant bacterial isolates showing higher accumulation of l-lactic acid belong to 3 groups of bacillus coagulans, according to their 16s rrna gene sequence similarities. b. coagulans strains m21 and m36 produced high amounts of l-lactic acid of high optical purity and lactic acid selectivity in model kitchen refuse medium and glucose-yeast extract ...201424119530
kinetic characterization of recombinant bacillus coagulans fdp-activated l-lactate dehydrogenase expressed in escherichia coli and its substrate specificity.bacillus coagulans is a homofermentative, acid-tolerant and thermophilic sporogenic lactic acid bacterium, which is capable of producing high yields of optically pure lactic acid. the l-(+)-lactate dehydrogenase (l-ldh) from b. coagulans is considered as an ideal biocatalyst for industrial production. in this study, the gene ldhl encoding a thermostable l-ldh was amplified from b. coagulans nl01 genomic dna and successfully expressed in escherichia coli bl21 (de3). the recombinant enzyme was par ...201424412354
draft genome sequence of bacillus coagulans gbi-30, 6086, a widely used spore-forming probiotic strain.bacillus coagulans gbi-30, 6086 is a safe strain, already available on the market, and characterized by certified beneficial effects. the draft genome sequence presented here constitutes the first pillar toward the identification of the molecular mechanisms responsible for its positive features and safety.201425377698
improving lactic acid productivity from wheat straw hydrolysates by membrane integrated repeated batch fermentation under non-sterilized conditions.bacillus coagulans ipe22 was used to produce lactic acid (la) from mixed sugar and wheat straw hydrolysates, respectively. all fermentations were conducted under non-sterilized conditions and sodium hydroxide was used as neutralizing agent to avoid the production of insoluble caso4. in order to eliminate the sequential utilization of mixed sugar and feedback inhibition during batch fermentation, membrane integrated repeated batch fermentation (mirb) was used to improve la productivity. with mirb ...201424811443
axone, an ethnic probiotic containing food, reduces age of sexual maturity and increases poultry production.axone (akhuni) is a homemade synbiotic (nagamese fermented soybean product) served as side dish in north eastern india. in this study, effects of axone feeding on growth, weight gain, sexual maturity and egg production on vanaraja birds (a strain of poultry bird developed at pdp hyderabad for villages and backyard poultry) were evaluated. axone incorporation in commercial poultry feed at the rate of 5% (w/w) significantly improved growth rate (weight gain) both in male (p 0.001) and female (p 0. ...201424801640
efficient calcium lactate production by fermentation coupled with crystallization-based in situ product removal.lactic acid is a platform chemical with various industrial applications, and its derivative, calcium lactate, is an important food additive. fermentation coupled with in situ product removal (ispr) can provide more outputs with high productivity. the method used in this study was based on calcium lactate crystallization. three cycles of crystallization were performed during the fermentation course using a bacillus coagulans strain h-1. as compared to fed-batch fermentation, this method showed 1. ...201424780270
predicting bacillus coagulans spores inactivation in tomato pulp under nonisothermal heat treatments.the knowledge and understanding of bacillus coagulans inactivation during a thermal treatment in tomato pulp, as well as the influence of temperature variation during thermal processes are essential for design, calculation, and optimization of the process. the aims of this work were to predict b. coagulans spores inactivation in tomato pulp under varying time-temperature profiles with gompertz-inspired inactivation model and to validate the model's predictions by comparing the predicted values w ...201424712665
an efficient process for lactic acid production from wheat straw by a newly isolated bacillus coagulans strain ipe22.a thermophilic lactic acid (la) producer was isolated and identified as bacillus coagulans strain ipe22. the strain showed remarkable capability to ferment pentose, hexose and cellobiose, and was also resistant to inhibitors from lignocellulosic hydrolysates. based on the strain's promising features, an efficient process was developed to produce la from wheat straw. the process consisted of biomass pretreatment by dilute sulfuric acid and subsequent sscf (simultaneous saccharification and co-fer ...201424679663
evaluation of high-intensity ultrasonication for the inactivation of endospores of 3 bacillus species in nonfat milk.endospores of bacillus licheniformis [american type culture collection (atcc) 6634], bacillus coagulans (atcc 12245), and geobacillus stearothermophilus (atcc 15952) were spiked in sterile nonfat milk, and subjected to high intensity batch ultrasonication treatment at different amplitudes (80 or 100%) and durations (1 to 10 min). increasing the amplitude from 80 to 100% did not result in enhanced inactivation of g. stearothermophilus endospores. however, an increase in the duration of ultrasonic ...201425087024
betaine and beet molasses enhance l-lactic acid production by bacillus coagulans.lactic acid is an important chemical with various industrial applications, and it can be efficiently produced by fermentation, in which bacillus coagulans strains present excellent performance. betaine can promote lactic acid fermentation as an effective osmoprotectant. here, positive effect of betaine on fermentation by b. coagulans is revealed. betaine could enhance lactic acid production by protecting l-ldh activity and cell growth from osmotic inhibition, especially under high glucose concen ...201424956474
enhanced l-lactic acid production from biomass-derived xylose by a mutant bacillus coagulans.xylose effective utilization is crucial for production of bulk chemicals from low-cost lignocellulosic substrates. in this study, an efficient l-lactate production process from xylose by a mutant bacillus coagulans nl-cc-17 was demonstrated. the nutritional requirements for l-lactate production by b. coagulans nl-cc-17 were optimized statistically in shake flask fermentations. corn steep liquor powder and yeast exact were identified as the most significant factors by the two-level plackett-burma ...201424879598
a randomized double-blind placebo-controlled clinical trial on efficacy and safety of association of simethicone and bacillus coagulans (colinox®) in patients with irritable bowel syndrome.irritable bowel syndrome (ibs) is a chronic gastrointestinal (gi) disorder that affects 15-20% of the western population.201424867512
analysis of the interaction between bacillus coagulans and bacillus thuringiensis s-layers and calcium ions by xrd, light microscopy, and ftir.s-layer is a self-assemble regularly crystalline surface that covers major cell wall component of many bacteria and archaea and exhibits a high metal-binding capacity. we have studied the effect of the calcium ions and type of solid support (glass or mica) on the structure of the s-layers from bacillus coagulans hn-68 and bacillus thuringiensis mh14 upon simple methods based on light microscopy and afm. furthermore, the fourier transform infrared spectroscopy (ftir) study is indicated that the c ...201424639088
genomic analysis of thermophilic bacillus coagulans strains: efficient producers for platform bio-chemicals.microbial strains with high substrate efficiency and excellent environmental tolerance are urgently needed for the production of platform bio-chemicals. bacillus coagulans has these merits; however, little genetic information is available about this species. here, we determined the genome sequences of five b. coagulans strains, and used a comparative genomic approach to reconstruct the central carbon metabolism of this species to explain their fermentation features. a novel xylose isomerase in t ...201424473268
effect of prebiotics on the fecal microbiota of elderly volunteers after dietary supplementation of bacillus coagulans gbi-30, 6086.in advancing age, gut populations of beneficial microbes, notably bifidobacterium spp., show a marked decline. this contributes to an environment less capable of maintaining homoeostasis. this in vitro investigation studied the possible synergistic effects of probiotic supplementation in modulating the gut microbiota enabling prebiotic therapy to in elderly persons. single stage batch culture anaerobic fermenters were used and inoculated with fecal microbiota obtained from volunteers after takin ...201425219857
major role of nad-dependent lactate dehydrogenases in the production of l-lactic acid with high optical purity by the thermophile bacillus coagulans.bacillus coagulans 2-6 is an excellent producer of optically pure l-lactic acid. however, little is known about the mechanism of synthesis of the highly optically pure l-lactic acid produced by this strain. three enzymes responsible for lactic acid production-nad-dependent l-lactate dehydrogenase (l-nldh; encoded by ldhl), nad-dependent d-lactate dehydrogenase (d-nldh; encoded by ldhd), and glycolate oxidase (gox)-were systematically investigated in order to study the relationship between these ...201425217009
a comparative proteomic analysis of bacillus coagulans in response to lactate stress during the production of l-lactic acid.the growth rate and maximum biomass of bacillus coagulans 2-6 were inhibited by lactate; inhibition by sodium lactate was stronger than by calcium lactate. the differences of protein expressions by b. coagulans 2-6 under the lactate stress were determined using two-dimensional electrophoresis coupled with mass spectrometric identification. under the non-stress condition, calcium lactate stress and sodium lactate stress, the number of detected protein spots was 1,571 ± 117, 1,281 ± 231 and 904 ±  ...201425214213
immunomodulation of antiretroviral drug-suppressed chronic hiv-1 infection in an oral probiotic double-blind placebo-controlled trial.a putative source of inappropriate immune activation that drives human immunodeficiency virus (hiv)-1 immunopathogenesis is the gastrointestinal tract. even with effective antiretroviral treatment, residual activation persists. we hypothesized that an oral probiotic could improve the residual immune activation in chronic treated hiv-1 infection, and tested a bacillus coagulans gbi-30, 6086 capsule probiotic in hiv-1-infected persons with suppressed viremia on stable antiretroviral therapy in a 3 ...201425127924
the efficacy of a synbiotic containing bacillus coagulans in treatment of irritable bowel syndrome: a randomized placebo-controlled trial.we aimed to evaluate the efficacy of a synbiotic containing bacillus coagulans in treatment of ibs.201425120896
dietary supplementation of probiotics affects growth, immune response and disease resistance of cyprinus carpio fry.the effects of dietary bacillus coagulans (mtcc 9872), bacillus licheniformis (mtcc 6824) and paenibacillus polymyxa (mtcc 122) supplementation on growth performance, non-specific immunity and protection against aeromonas hydrophila infection were evaluated in common carp, cyprinus carpio fry. laboratory maintained b. coagulans, b. licheniformis and p. polymyxa were used to study antagonistic activity against fish pathogenic bacteria by agar well diffusion assay. healthy fish fry were challenged ...201425160796
inactivation of bacillus spores inoculated in milk by ultra high pressure homogenization.ultra high-pressure homogenization treatments at 300 mpa with inlet temperatures (ti) of 55, 65, 75 and 85 °c were applied to commercial ultra high temperature treated whole milk inoculated with bacillus cereus, bacillus licheniformis, bacillus sporothermodurans, bacillus coagulans, geobacillus stearothermophilus and bacillus subtilis spores in order to evaluate the inactivation level achieved. ultra high-pressure homogenization conditions at 300 mpa with ti = 75 and 85 °c were capable of a spor ...201425084664
bacillus species isolated from tungrymbai and bekang, naturally fermented soybean foods of india.tungrymbai and bekang are naturally fermented soybean foods commonly consumed in meghalaya and mizoram states of india. a total of 39 samples of tungrymbai and 43 samples of bekang were collected from different villages and markets of meghalaya and mizoram, respectively and were analysed for microbial load. in both tungrymbai and bekang, the average population of bacillus spp. was 8.2±0.1 log cfu/g. a total of 428 isolates of bacillus were isolated from tungrymbai (211) and bekang (217) for deta ...201525574846
[composition diversity and metabolic characters of lactic acid bacteria community sgl].we aimed to select a stable lactic acid bacteria community from switchgrass silage, that was efficient in lactic acid production.201526915229
differential oxidative stress responses in castor semilooper, achaea janata.balance between reactive oxygen species (ros) and the antioxidant (ao) defense mechanisms is vital for organism survival. insects serve as an ideal model to elucidate oxidative stress responses as they are prone to different kinds of stress during their life cycle. the present study demonstrates the modulation of ao enzyme gene expression in the insect pest, achaea janata (castor semilooper), when subjected to different oxidative stress stimuli. antioxidant enzymes' (catalase (cat), superoxide d ...201526455997
purification and characterization of a novel plantaricin, kl-1y, from lactobacillus plantarum kl-1.three bacteriocins from lactobacillus plantarum kl-1 were successfully purified using ammonium sulfate precipitation, cation-exchange chromatography and reverse-phase hplc. the bacteriocin peptides kl-1x, -1y and -1z had molecular masses of 3053.82, 3498.16 and 3533.16 da, respectively. all three peptides were stable at ph 2-12 and 25 °c and at high temperatures of 80 and 100 °c for 30 min and 121 °c for 15 min. however, they differed in their susceptibility to proteolytic enzymes and their inhi ...201525862353
biochemical properties and yields of diverse bacterial laccase-like multicopper oxidases expressed in escherichia coli.laccases are multi-copper oxidases that oxidize a broad range of substrates at the expense of molecular oxygen, without any need for co-factor regeneration. these enzymes bear high potential for the sustainable synthesis of fine chemicals and the modification of (bio)polymers. here we describe cloning and expression of five novel bacterial laccase-like multi copper oxidases (lmcos) of diverse origin which were identified by homology searches in online databases. activity yields under different e ...201526068013
the effects of probiotic, prebiotic and synbiotic diets containing bacillus coagulans and inulin on rat intestinal microbiota.an in vivo experiment was conducted to study the effects of probiotic bacillus coagulans spores, with and without prebiotic, inulin, on gastrointestinal (gi) microbiota of healthy rats and its potentiality to survive in the gi tract. forty-eight male wistar rats were randomly divided into four groups (n=12) and fed as follows: standard diet (control), standard diet supplied with 5% w/w long chain inulin (prebiotic), standard diet with 10(9)/day spores of b. coagulans by orogastric gavage (probio ...201527175187
draft genome sequence of bacillus coagulans nl01, a wonderful l-lactic acid producer.here, we report the draft genome sequence of bacillus coagulans nl01, which could produce high optically pure l-lactic acid using xylose as a sole carbon source. the draft genome is 3,505,081 bp, with 144 contigs. about 3,903 protein-coding genes and 92 rrnas are predicted from this assembly.201526089419
bacteriocinogenic potential of a probiotic strain bacillus coagulans [bdu3] from ngari.bacteriocin producing strain bdu3 was isolated from a traditional fermented fish of manipur ngari. the strain bdu3 was identified as bacillus coagulans by phenotypic and genotypic characterization. the bdu3 produced novel bacteriocin, which showed an antimicrobial spectrum toward a wide spectrum of food borne, and closely related pathogens with a mic that ranged between 0.5 and 2.5 μg/ml. the isolate was able to tolerate ph as low as 2.0 and up to 0.2% bile salt concentration. three step purific ...201526054664
comparison of the effects of ph-dependent peppermint oil and synbiotic lactol (bacillus coagulans + fructooligosaccharides) on childhood functional abdominal pain: a randomized placebo-controlled study.still there is no consensus on the best treatment for abdominal pain-related functional gastrointestinal disorders (fgids).201526023339
production of optically pure l-phenyllactic acid by using engineered escherichia coli coexpressing l-lactate dehydrogenase and formate dehydrogenase.l-phenyllactic acid (l-pla) is a novel antiseptic agent with broad and effective antimicrobial activity. in addition, l-pla has been used for synthesis of poly(phenyllactic acid)s, which exhibits better mechanical properties than poly(lactic acid)s. however, the concentration and optical purity of l-pla produced by native microbes was rather low. an nad-dependent l-lactate dehydrogenase (l-nldh) from bacillus coagulans nl01 was confirmed to have a good ability to produce l-pla from phenylpyruvic ...201526008622
genome sequence of bacillus coagulans p38, an efficient polymer-grade l-lactate producer from cellulosic substrates.bacillus coagulans p38 is an efficient polymer-grade l-lactic acid producer from a cellulosic carbon source. here, the draft 3.37-mb genome sequence of this potential strain may provide useful information to further improve the strain performance for higher titers and, importantly, to understand the mechanism of its high tolerance for 2-furfural.201525999580
the effect of bacillus coagulans-fermented and nonfermented ginkgo biloba on the immunity status of broiler chickens.to evaluate and compare the effects of bacillus coagulans-fermented ginkgo biloba (fg) and nonfermented ginkgo biloba (nfg) on the immunity status of broiler chickens, 180 1-d-old female arbor acres chicks were divided into 3 groups and fed either a basal diet, a basal diet supplemented with 0.3% nfg, or a basal diet supplemented with 0.3% fg. blood samples were taken on the seventh (before vaccination), 14th, 21st, 28th and 35th day for the assessment of serum il-18 and interferon γ (ifn-γ) lev ...201526440007
potential probiotic attributes of a new strain of bacillus coagulans cgmcc 9951 isolated from healthy piglet feces.a new strain of bacillus coagulans cgmcc 9551, which has a broad range of antibacterial activities against six main pathogenic bacteria including escherichia coli o8, staphylococcus aureus, salmonella enterica subsp. enterica serovar enteritidis, streptococcus suis, listeria monocytogenes and pasteurella multocida, was isolated from healthy piglet feces. in adhesion assay, the isolate exhibited a stronger adhesion to pig intestinal mucus than that of b. subtilis jt143 and l. acidophilus ly24 res ...201525752235
high-titer lactic acid production from naoh-pretreated corn stover by bacillus coagulans la204 using fed-batch simultaneous saccharification and fermentation under non-sterile condition.lactic acid (la) is an important chemical with various industrial applications. non-food feedstock is commercially attractive for use in la production; however, efficient la fermentation from lignocellulosic biomass resulting in both high yield and titer faces technical obstacles. in this study, the thermophilic bacterium bacillus coagulans la204 demonstrated considerable ability to ferment glucose, xylose, and cellobiose to la. importantly, la204 produces la from several naoh-pretreated agro st ...201525704098
synbiotic containing bacillus coagulans and fructo-oligosaccharides for functional abdominal pain in children.we evaluated the effectiveness of a synbiotic in the treatment of childhood functional abdominal pain (fap).201525584177
augmented biogas production from protein-rich substrates and associated metagenomic changes.this study demonstrates that appropriate adaptation of the microbial community to protein-rich biomass can lead to sustainable biogas production. the process of acclimation to these unusual mono-substrates was controlled by the protease activity of the microbial community. meat extract (c/n=3.32) and kitchen waste (c/n=12.43) were used as biogas substrates. metagenome analysis highlighted several mesophilic strains that displayed a preference for protein degradation. bacillus coagulans, bacillus ...201525316194
enhanced pc12 cells proliferation with self-assembled s-layer proteins scaffolds.finding 3d biocompatible and biodegradable scaffold is important in tissue engineering which plays a critical role in transplanting methods. several biomaterials, such as poly-l-lactide, poly(lactic-co-glycolic acid), poly(l-lactic acid)/poly(lactide-co-glycolide), alginate, collagen gel, and so on, have been applied as scaffold to culture cells in 3d environment. the most significant problem of the synthetic materials is lack of biocompatibility and bioactivity. herein, self assemble s-layer pr ...201525253266
open fermentative production of l-lactic acid using white rice bran by simultaneous saccharification and fermentation.to reduce raw material cost for lactic acid production, white rice bran as an important byproduct in rice milling, was used in l-lactic acid production by open simultaneous saccharification and fermentation (ssf). although one thermotolerant strain was used at a temperature as high as 50°c, the open fermentation was still inefficient due to the indigenous thermophilic bacteria from corn steep liquor powder. a stepwise controlled ph was proposed in open ssf process, and no complicated pretreatmen ...201526433792
isolation and identification of poly β-hydroxybutyrate over-producing bacteria and optimization of production medium.biodegradable polyesters are candidates for the development of environmental friendly plastics. poly-β-hydroxybutyrate (phb) is a type of polyester from the hydroxyalkanoates family, synthesized by bacteria as an intracellular material and accumulated as granules in the cytoplasm.201526396711
microbial production of lactic acid: the latest development.lactic acid is an important platform chemical for producing polylactic acid (pla) and other value-added products. it is naturally produced by a wide spectrum of microbes including bacteria, yeast and filamentous fungi. in general, bacteria ferment c5 and c6 sugars to lactic acid by either homo- or hetero-fermentative mode. xylose isomerase, phosphoketolase, transaldolase, l- and d-lactate dehydrogenases are the key enzymes that affect the ways of lactic acid production. metabolic engineering of ...201526260005
efficient hydrolysis of corncob residue through cellulolytic enzymes from trichoderma strain g26 and l-lactic acid preparation with the hydrolysate.to prepare fermentable hydrolysate from corncob residue (ccr), trichoderma strain g26 was cultured on medium containing ccr for production of cellulolytic enzymes through solid-state fermentation (ssf), resulting in 71.3 iu/g (fpa), 136.2 iu/g (cmcase), 85.1 iu/g (β-glucosidase) and 11,344 iu/g (xylanase), respectively. through a three-stage saccharification strategy, ccr was hydrolyzed by the enzymatic solution (6.5 fpu/ml) into fermentable hydrolysate containing 60.1g/l glucose (81.2% cellulos ...201526143000
utilization of coconut oil cake for the production of lipase using bacillus coagulans vkl1.the overproduction of enzymes was performed by manipulating the medium components. in our study, solvent-tolerant thermophilic lipase-producing bacillus coagulans was isolated from soil samples and a stepwise optimization strategy was employed to increase the lipase production using coconut oil cake basal medium. in the first step, the influence of ph, temperature, carbon source, nitrogen source and inducers on lipase activity was investigated by the one-factor-at-a-time (ofat) method. in the se ...201526133510
hemp hurds biorefining: a path to green l-(+)-lactic acid production.sugars streams generated by organosolv pretreatment of hemp hurds, cellulose (c6) and hemicellulose (c5) fractions, were fermented to lactic acid (la) by bacillus coagulans strains xzl4 and dsm1. pretreatment conditions and enzymatic hydrolysis were optimized and b. coagulans aptness to use lignocellulosic-derived sugars as a carbon source was evaluated. methanolic organosolv pretreatment with 2.5% (w/w) h2so4 gave the best results in terms of glucan recovery (98%), enzymatic hydrolysis of pretr ...201525983223
combined pressure-thermal inactivation effect on spores in lu-wei beef--a traditional chinese meat product.this study investigated the inactivation effect and kinetics of bacillus coagulans and geobacillus stearothermophilus spores suspended in lu-wei beef by combining high pressure (500 and 600 mpa) and moderate heat (70 and 80 °c or 80 and 90 °c).201525963751
bacillus coagulans gbi-30, 6086 modulates faecalibacterium prausnitzii in older men and women.advancing age is linked to a decrease in beneficial bacteria such as bifidobacterium spp. and reduced aspects of innate immune function.201525948780
comparative transcriptome analysis reveals different molecular mechanisms of bacillus coagulans 2-6 response to sodium lactate and calcium lactate during lactic acid production.lactate production is enhanced by adding calcium carbonate or sodium hydroxide during fermentation. however, bacillus coagulans 2-6 can produce more than 180 g/l l-lactic acid when calcium lactate is accumulated, but less than 120 g/l l-lactic acid when sodium lactate is formed. the molecular mechanisms by which b. coagulans responds to calcium lactate and sodium lactate remain unclear. in this study, comparative transcriptomic methods based on high-throughput rna sequencing were applied to stud ...201525875592
effects of dietary fiber with bacillus coagulans lilac-01 on bowel movement and fecal properties of healthy volunteers with a tendency for constipation.to elucidate the effects of lilac lab (bacillus coagulans lilac-01 and okara [soy pulp] powder) on bowel movements/fecal properties, we conducted a double-blind placebo-controlled randomized trial with healthy japanese volunteers with a tendency for constipation (n = 297). the subjects ingested 2 g/d placebo (okara powder) or test food (lilac lab, 1 × 10(8) cfu) once a day for 2 weeks. in the test group of functionally constipated subjects, the changes in the average scores of self-reported feca ...201525338680
microbial production of lactic acid: the latest development.lactic acid is an important platform chemical for producing polylactic acid (pla) and other value-added products. it is naturally produced by a wide spectrum of microbes including bacteria, yeast and filamentous fungi. in general, bacteria ferment c5 and c6 sugars to lactic acid by either homo- or hetero-fermentative mode. xylose isomerase, phosphoketolase, transaldolase, l- and d-lactate dehydrogenases are the key enzymes that affect the ways of lactic acid production. metabolic engineering of ...201626287368
probiotic bacillus coagulans gbi-30, 6086 reduces exercise-induced muscle damage and increases recovery.objective. probiotics have been reported to support healthy digestive and immune function, aid in protein absorption, and decrease inflammation. further, a trend to increase vertical jump power has been observed following co-administration of protein and probiotics in resistance-trained subjects. however, to date the potential beneficial effect of probiotics on recovery from high intensity resistance exercise have yet to be explored. therefore, this study examined the effect of co-administration ...201627547577
effect of oral administration of bacillus coagulans b37 and bacillus pumilus b9 strains on fecal coliforms, lactobacillus and bacillus spp. in rat animal model.to investigate the effect of oral administration of two bacillus strains on fecal coliforms, lactobacillus and bacillus spp. in rat animal model.201627536040
highly efficient production of optically pure l-lactic acid from corn stover hydrolysate by thermophilic bacillus coagulans.a thermophilic strain bacillus coagulans (nbrc 12714) was employed to produce l-lactic acid from corn stover hydrolysate in membrane integrated continuous fermentation. the strain nbrc 12714 metabolized glucose and xylose by the embden-meyerhof-parnas pathway (emp) and the pentose phosphate pathway (ppp), producing l-lactic acid with optical purity >99.5%. the overall l-lactic acid titer of 92g/l with a yield of 0.91g/g and a productivity of 13.8g/l/h were achieved at a dilution rate of 0.15h(-1 ...201627479802
the optimization of l-lactic acid production from sweet sorghum juice by mixed fermentation of bacillus coagulans and lactobacillus rhamnosus under unsterile conditions.the cost reduction of raw material and sterilization could increase the economic feasibility of l-lactic acid fermentation, and the development of an cost-effective and efficient process is highly desired. to improve the efficiency of open fermentation by lactobacillus rhamnosus based on sweet sorghum juice (ssj) and to overcome sucrose utilization deficiency of bacillus coagulans, a mixed fermentation was developed. besides, the optimization of ph, sugar concentration and fermentation medium we ...201627469090
the effects of orally administered bacillus coagulans and inulin on prevention and progression of rheumatoid arthritis in rats.probiotics have been considered as an approach to addressing the consequences of different inflammatory disorders. the spore-forming probiotic strain bacillus coagulans has demonstrated anti-inflammatory and immune-modulating effects in both animals and humans. the prebiotic inulin also potentially affects the immune system as a result of the change in the composition or fermentation profile of the gastrointestinal microbiota.201627427194
two highly adhesive lactic acid bacteria strains are protective in zebrafish infected with aeromonas hydrophila by evocation of gut mucosal immunity.to increase the knowledge of probiotic effects and potential mechanisms, we report on the use of the zebrafish model to investigate the in vivo colonization ability, as well as the protective effects associated with gut mucosal immune barrier and responses against aeromonas hydrophila infection of previously characterized probiotic lactic acid bacteria (lab) strains, bacillus coagulans 09·712 and lactobacillus plantarum 08·923, in comparison with that of three commercialized strains.201626555459
precultivation of bacillus coagulans dsm2314 in the presence of furfural decreases inhibitory effects of lignocellulosic by-products during l(+)-lactic acid fermentation.by-products resulting from thermo-chemical pretreatment of lignocellulose can inhibit fermentation of lignocellulosic sugars to lactic acid. furfural is such a by-product, which is formed during acid pretreatment of lignocellulose. ph-controlled fermentations with 1 l starting volume, containing yp medium and a mixture of lignocellulosic by-products, were inoculated with precultures of bacillus coagulans dsm2314 to which 1 g/l furfural was added. the addition of furfural to precultures resulted ...201627464829
combination of soya pulp and bacillus coagulans lilac-01 improves intestinal bile acid metabolism without impairing the effects of prebiotics in rats fed a cholic acid-supplemented diet.intestinal bacteria are involved in bile acid (ba) deconjugation and/or dehydroxylation and are responsible for the production of secondary ba. however, an increase in the production of secondary ba modulates the intestinal microbiota due to the bactericidal effects and promotes cancer risk in the liver and colon. the ingestion of bacillus coagulans improves constipation via the activation of bowel movement to promote defaecation in humans, which may alter ba metabolism in the intestinal content ...201627464459
high-throughput system for screening of high l-lactic acid-productivity strains in deep-well microtiter plates.for strain improvement, robust and scalable high-throughput cultivation systems as well as simple and rapid high-throughput detection methods are crucial. however, most of the screening methods for lactic acid bacteria (lab) strains were conducted in shake flasks and detected by high-performance liquid chromatography (hplc), making the screening program laborious, time-consuming and costly. in this study, an integrated strategy for high-throughput screening of high l-lactic acid-productivity str ...201627447955
physicochemical, microbiological and spoilage analysis of probiotic processed cheese analogues with reduced emulsifying salts during refrigerated storage.microbial quality of low-salt processed cheeses supplemented with bacillus coagulans spores (10(7)-10(8) cfu/g) relying on their physicochemical characteristics during 60 day-cold storage was evaluated. a reduction in moisture content, water activity and ph value and a significant enhancement in proteolytic index of control and probiotic samples were obtained by prolonging storage time. survival rate of the probiotic cells significantly decreased up to day 30, while total count of the viable cel ...201627162379
complete genome sequence of probiotic bacillus coagulans hm-08: a potential lactic acid producer.bacillus coagulans hm-08 is a commercialized probiotic strain in china. its genome contains a 3.62mb circular chromosome with an average gc content of 46.3%. in silico analysis revealed the presence of one xyl operon as well as several other genes that are correlated to xylose utilization. the genetic information provided here may help to expand its future biotechnology potential in lactic acid production.201627130497
draft genome sequence of the spore-forming probiotic strain bacillus coagulans unique is-2.italic! bacillus coagulansunique is-2 is a potential spore-forming probiotic that is commercially available on the market. the draft genome sequence presented here provides deep insight into the beneficial features of this strain for its safe use as a probiotic for various human and animal health applications.201627103709
fermentative utilization of coffee mucilage using bacillus coagulans and investigation of down-stream processing of fermentation broth for optically pure l(+)-lactic acid production.in this study, mucilage, a residue from coffee production, was investigated as substrate in fermentative l(+)-lactic acid production. mucilage was provided as liquid suspension consisting glucose, galactose, fructose, xylose and sucrose as free sugars (up to 60gl(-1)), and used directly as medium in bacillus coagulans batch fermentations carried out at 2 and 50l scales. using mucilage and 5gl(-1) yeast extract as additional nitrogen source, more than 40gl(-1) lactic acid was obtained. productivi ...201627035470
an innovative optical sensor for the online monitoring and control of biomass concentration in a membrane bioreactor system for lactic acid production.accurate real-time process control is necessary to increase process efficiency, and optical sensors offer a competitive solution because they provide diverse system information in a noninvasive manner. we used an innovative scattered light sensor for the online monitoring of biomass during lactic acid production in a membrane bioreactor system because biomass determines productivity in this type of process. the upper limit of the measurement range in fermentation broth containing bacillus coagul ...201627007380
statistical optimization for enhanced yields of probiotic bacillus coagulans and its phage resistant mutants followed by kinetic modelling of the process.probiotics are microorganisms which when administered in adequate amounts confer health benefits to the host. a leading pharmaceutical company producing bacillus coagulans as a probiotic was facing the problem of recurring phage attacks. two mutants viz. b. co piii and b. co miii that were isolated as phage resistant mutants after uv irradiation and mms treatment of phage sensitive b. coagulans parental culture were characterized at functional and molecular level and were noted to have undergone ...201627722071
antibacterial effect of fructose laurate synthesized by candida antarctica b lipase-mediated transesterification.sugar esters are valuable compounds composed of various sugars and fatty acids that can be used as antibacterial agents and emulsifiers in toothpaste and canned foods. for example, fructose fatty acid esters suppress growth of streptococcus mutans, a typical pathogenic bacterium causing dental caries. in this study, fructose laurate ester was chosen as a target material and was synthesized by a transesterification reaction using candida antarctica lipase b. we performed a solvent screening exper ...201627291679
systematic development and optimization of chemically defined medium supporting high cell density growth of bacillus coagulans.with determined components and experimental reducibility, the chemically defined medium (cdm) and the minimal chemically defined medium (mcdm) are used in many metabolism and regulation studies. this research aimed to develop the chemically defined medium supporting high cell density growth of bacillus coagulans, which is a promising producer of lactic acid and other bio-chemicals. in this study, a systematic methodology combining the experimental technique with flux balance analysis (fba) was p ...201627262567
complete genomes of bacillus coagulans s-lac and bacillus subtilis to-a jpc, two phylogenetically distinct probiotics.several spore-forming strains of bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. we report the complete genomes of two commercially available probiotics, bacillus coagulans s-lac and bacillus subtilis to-a jpc, and compare them with the genomes of other bacillus and lactobacillus. the taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping pro ...201627258038
integrate genome-based assessment of safety for probiotic strains: bacillus coagulans gbi-30, 6086 as a case study.probiotics are microorganisms that confer beneficial effects on the host; nevertheless, before being allowed for human consumption, their safety must be verified with accurate protocols. in the genomic era, such procedures should take into account the genomic-based approaches. this study aims at assessing the safety traits of bacillus coagulans gbi-30, 6086 integrating the most updated genomics-based procedures and conventional phenotypic assays. special attention was paid to putative virulence ...201626952108
evaluation of genetic and phenotypic consistency of bacillus coagulans mtcc 5856: a commercial probiotic strain.commercial probiotics preparation containing bacillus coagulans have been sold in the market for several decades. due to its high intra-species genomic diversity, it is very likely that b. coagulans strain may alter in different ways over multiple years of production. therefore, the present study focuses to evaluate the genetic consistency and probiotic potential of b. coagulans mtcc 5856. phenotypic and genotypic techniques including biochemical profiling, 16s rrna sequencing, gtg 5″, box pcr f ...201626925622
bacillus coagulans mtcc 5856 supplementation in the management of diarrhea predominant irritable bowel syndrome: a double blind randomized placebo controlled pilot clinical study.bacillus coagulans mtcc 5856 has been marketed as a dietary ingredient, but its efficacy in diarrhea predominant irritable bowel syndrome (ibs) condition has not been clinically elucidated till date. thus, a double blind placebo controlled multi-centered trial was planned to evaluate the safety and efficacy of b. coagulans mtcc 5856 in diarrhea predominant ibs patients.201626922379
contributory roles of two l-lactate dehydrogenases for l-lactic acid production in thermotolerant bacillus coagulans.thermotolerant bacillus coagulans is considered to be a more promising producer for bio-chemicals, due to its capacity to withstand harsh conditions. two l-lactate dehydrogenase (ldh) encoding genes (ldhl1 and ldhl2) and one d-ldh encoding gene (ldhd) were annotated from the b. coagulans dsm1 genome. transcriptional analysis revealed that the expression of ldhl2 was undetectable while the ldhl1 transcription level was much higher than that of ldhd at all growth phases. deletion of the ldhl2 gene ...201627885267
genome sequences of 12 spore-forming bacillus species, comprising bacillus coagulans, bacillus licheniformis, bacillus amyloliquefaciens, bacillus sporothermodurans, and bacillus vallismortis, isolated from foods.here, we report the draft genomes of twelve isolates of five different bacillus species, all spore-forming, gram-positive bacteria.201627174261
diammonium phosphate stimulates transcription of l-lactate dehydrogenase leading to increased l-lactate production in the thermotolerant bacillus coagulans strain.exploration of cost-effective fermentation substrates for efficient lactate production is an important economic objective. although some organic nitrogen sources are also cheaper, inorganic nitrogen salts for lactate fermentation have additional advantages in facilitating downstream procedures and significantly improving the commercial competitiveness of lactate production. in this study, we first established an application of diammonium phosphate to replace yeast extract with a reduced 90 % nit ...201626883345
simultaneous production of l-lactic acid with high optical activity and a soil amendment with food waste that demonstrates plant growth promoting activity.a unique method to produce highly optically-active l-lactic acid and soil amendments that promote plant growth from food waste was proposed. three bacillus strains bacillus subtilis kbku21, b. subtilis n3-9 and bacillus coagulans t27, were used. strain kbku21 accumulated 36.9 g/l l-lactic acid with 95.7% optical activity and 98.2% l-lactic acid selectivity when fermented at 43°c for 84 h in a model kitchen refuse (mkr) medium. residual precipitate fraction (anaerobically-fermented mkr (afm) comp ...201626819060
bacterial nanocellulose-pectin bionanocomposites as prebiotics against drying and gastrointestinal condition.various encapsulating materials have been suggested to protect probiotics, but the potential of nanomaterials is yet to be exploited. this study aimed to improve the survivability of bacillus coagulans entrapping into bionanocomposites comprising of bacterial nanocellulose (bnc), pectin and schizophyllum commune extract were investigated as new matrices to protect probiotics. the bionanocomposite design was optimized to obtain the highest prebiotic score and survivability of probiotic under dryi ...201626627598
fermentative production of enantiomerically pure s-1,2-propanediol from glucose by engineered e. coli strain.the pure stereoisomers of 1,2-propanediol (1,2-pdo) could be used as starting materials to synthesize high value-added specialty chemicals and chiral pharmaceutical products. as the stereoisomers of 1,2-pdo cannot be obtained by traditional chemical synthesis processes, biotechnological processes have gained increasing attention. however, to our knowledge, the production of s-1,2-pdo directly from glucose has not been previously reported. in this study, we demonstrate a novel artificial pathway ...201626454866
cost-effective simultaneous saccharification and fermentation of l-lactic acid from bagasse sulfite pulp by bacillus coagulans cc17.the main barriers to cost-effective lactic acid production from lignocellulose are the high cost of enzymes and the ineffective utilization of the xylose within the hydrolysate. in the present study, the thermophilic bacillus coagulans strain cc17 was used for the simultaneous saccharification and fermentation (ssf) of bagasse sulfite pulp (bsp) to produce l-lactic acid. unexpectedly, ssf by cc17 required approximately 33.33% less fungal cellulase than did separate hydrolysis and fermentation (s ...201627750196
effect of low ph start-up on continuous mixed-culture lactic acid fermentation of dairy effluent.mixed-culture fermentation that does not require an energy-intensive sterilization process is a viable approach for the economically feasible production of lactic acid (la) due to the potential use of organic waste as feedstock. this study investigated mixed-culture la fermentation of whey, a high-strength organic wastewater, in continuous mode. variations in the hydraulic retention time (hrt) from 120 to 8 h under different ph regimes in two thermophilic reactors (55 °c) were compared for their ...201627709285
novel ph control strategy for efficient production of optically active l-lactic acid from kitchen refuse using a mixed culture system.uninvestigated control factors of meta-fermentation, the fermentative production of pure chemicals and fuels in a mixed culture system, were examined for production of optically pure l-lactic acid (la) from food waste. in meta-fermentations by ph swing control, l-la production with 100% optical purity (opl-la) was achieved even using unsterilized model kitchen refuse medium with preferential proliferation of l-la-producing bacillus coagulans, a minor member in the seed, whereas agitation decreas ...201627233097
genomic analysis of a xylose operon and characterization of novel xylose isomerase and xylulokinase from bacillus coagulans nl01.to investigate the xylose operon and properties of xylose isomerase and xylulokinase in bacillus coagulans that can effectively ferment xylose to lactic acid.201627206341
production of high concentration of l-lactic acid from cellobiose by thermophilic bacillus coagulans wcp10-4.thermophilic bacillus coagulans wcp10-4 is found to be able to convert cellobiose to optically pure l-lactic acid. its β-glucosidase activity is detected in whole cells (7.3 u/g dry cells) but not in culture medium, indicating the intracellular location of the enzyme. its β-glucosidase activity is observed only when cultured using cellobiose as the sole carbon source, indicating that the expression of this enzyme is tightly regulated in cells. the enzyme is most active at 50 °c and ph 7.0. the s ...201627183994
production of l(+)-lactic acid from acid pretreated sugarcane bagasse using bacillus coagulans dsm2314 in a simultaneous saccharification and fermentation strategy.sugars derived from lignocellulose-rich sugarcane bagasse can be used as feedstock for production of l(+)-lactic acid, a precursor for renewable bioplastics. in our research, acid-pretreated bagasse was hydrolysed with the enzyme cocktail gc220 and fermented by the moderate thermophilic bacterium bacillus coagulans dsm2314. saccharification and fermentation were performed simultaneously (ssf), adding acid-pretreated bagasse either in one batch or in two stages. ssf was performed at low enzyme do ...201627872661
comparison of high-titer lactic acid fermentation from naoh- and nh3-h2o2-pretreated corncob by bacillus coagulans using simultaneous saccharification and fermentation.lignocellulose is one of the most abundant renewable feedstocks that has attracted considerable attention as a substrate for biofuel and biochemical production. one such biochemical product, lactic acid, is an important fermentation product because of its great potential for the production of biodegradable and biocompatible polylactic acid. high-titer lactic acid production from lignocellulosic materials has been achieved recently; however, it requires biodetoxification or results in large amoun ...201627853308
relationship between the peroxidation of leukocytes index ratio and the improvement of postprandial metabolic stress by a functional food.for the first time, we investigated the relationship between postprandial dysmetabolism and the peroxidation of leukocytes index ratio (plir), a test that measures the resistance of leukocytes to exogenous oxidative stress and their functional capacity of oxidative burst upon activation. following a blind, placebo controlled, randomized, crossover design, ten healthy subjects ingested, in two different occasions, a high fat and high carbohydrates meal with snello cookie (hfhcm-s) or with control ...201626962396
effect of bacillus subtilis natto on meat quality and skatole content in topigs pigs.this study investigated the effect of bacillus subtilis (b. subtilis) natto on meat quality and skatole in topigs pigs. sixty topigs pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen) and fed with basic diet (control group), basic diet plus 0.1% b. subtilis natto (b group), and basic diet plus 0.1% b. subtilis natto plus 0.1% b. coagulans (bb group), respectively. all pigs were sacrificed at 100 kg. growth performance, meat quality, serum parameters and ...201626954164
leguminose green juice as an efficient nutrient for l(+)-lactic acid production.lactic acid is one of the most important building blocks for the production of bioplastic. many investigations have been conducted to reduce the lactic acid production costs. in this work, the focus was put on the application of legume pressed juice or green juice as nutrient source. the pressed juice was utilized directly without prior pre-treatment and sterilization. using two different alfalfa green juices and a clover green juice from two different harvest years as sole nutrients, non-steril ...201627422353
characterization of an l-arabinose isomerase from bacillus coagulans nl01 and its application for d-tagatose production.l-arabinose isomerase (ai) is a crucial catalyst for the biotransformation of d-galactose to d-tagatose. in previous reports, ais from thermophilic bacterial strains had been wildly researched, but the browning reaction and by-products formed at high temperatures restricted their applications. by contrast, ais from mesophilic bacillus strains have some different features including lower optimal temperatures and lower requirements of metallic cofactors. these characters will be beneficial to the ...201627363468
fermentative lactic acid production from coffee pulp hydrolysate using bacillus coagulans at laboratory and pilot scales.in this study, the lignocellulosic residue coffee pulp was used as carbon source in fermentative l(+)-lactic acid production using bacillus coagulans. after thermo-chemical treatment at 121°c for 30min in presence of 0.18moll(-1) h2so4 and following an enzymatic digestion using accellerase 1500 carbon-rich hydrolysates were obtained. two different coffee pulp materials with comparable biomass composition were used, but sugar concentrations in hydrolysates showed variations. the primary sugars we ...201627359065
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