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Probiotics: Shaping the gut immunological responses

World Journal of Gastroenterology 2024年第15期30卷 2096-2108页

作者： Eirini Filidou Leonidas Kandilogiannakis Anne Shrewsbury George Kolios Katerina Kotzampassi Faculty of Medicine Laboratory of PharmacologyDemocritus University of ThraceAlexandroupolis 68100Greece Department of Surgery Aristotle University of ThessalonikiThessaloniki 54636Greece

Probiotics are live microorganisms exerting beneficial effects on the host’s health when administered in adequate amounts.Among the most popular and adequately studied probiotics are bacteria from the families Lactobacillaceae,Bifidobacteriaceae and yeasts.Most of them have been shown,both in vitro and in vivo studies of intestinal inflammation models,to provide favorable results by means of improving the gut microbiota composition,promoting the wound healing process and shaping the immunological responses.Chronic intestinal conditions,such as inflammatory bowel diseases(IBD),are characterized by an imbalance in microbiota composition,with decreased diversity,and by relapsing and persisting inflammation,which may lead to mucosal damage.Although the results of the clinical studies investigating the effect of probiotics on patients with IBD are still controversial,it is without doubt that these microorganisms and their metabolites,now named postbiotics,have a positive influence on both the host’s microbiota and the immune system,and ultimately alter the topical tissue microenvironment.This influence is achieved through three axes:(1)By dis-placement of potential pathogens via competitive exclusion;(2)by offering protection to the host through the secretion of various defensive mediators;and(3)by supplying the host with essential nutrients.We will analyze and discuss almost all the in vitro and in vivo studies of the past 2 years dealing with the possible favorable effects of certain probiotic genus on gut immunological responses,highlighting which species are the most beneficial against intestinal inflammation.

关键词： Probiotics Lactobacillaceae Bifidobacteriaceae saccharomyces Intestinal inflammation Immune responses

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Effect of saccharomyces boulardii and Bacillus subtilis B10 on gut microbiota modulation in broilers

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Animal Nutrition 2018年第4期4卷 358-366页

作者： Chubin Qin Li Gong Xiaoping Zhang Yuanyuan Wang Yibin Wang Baikui Wang Yali Li Weifen Li Key Laboratory of Molecular Animal Nutrition Ministry of Education College of Animal Science Zhejiang University China National Bamboo Research Center Key Laboratory of High Efficient Processing of Bamboo of Zhejiang Province

The gut microbiota plays important roles in animal overall health and productiveness. Balancing host gut microbiota by probiotics has been documented. Our previous study showed that saccharomyces boulardii(Sb) and Bacillus subtilis B10(Bs) significantly improve growth performance and modulate the intestinal histomorphology in broilers. To increase the knowledge regarding Sb and Bs, this study investigated the effects of these 2 probiotic strains on the gut microbiota in broilers. Three hundred 1-day-old Sanhuang broilers(Chinese cross breed) were randomly divided into 3 groups, each group with 5 replications(n = 20). The control group(CK) was fed a basal diet containing an antibiotic(virginiamycin, 20 mg/kg) and the other 2 groups received Sb and Bs(1 × 10~8cfu/kg of feed) in addition to the basal diet. After 72 d of treatment, pyrosequencing revealed that the bacterial communities varied along the section of intestinal tract in the control and Bs groups, but not in the Sb group. No difference in microbial diversity was observed among 3 groups. The major phyla observed along the GI tract of broilers(particularly in the duodenum and cecum) were Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobia, which were considered potentially growth performance-related. Bacteroidetes, Proteobacteria, and Verrucomicrobia were observed at a much higher abundance in the jejunums and ileums of the Sb group(P < 0.05). In addition, the jejunal microbial communities formed 3 different clusters at either the genus level or the category of metabolism among the groups, based on the principal component analyses. These data indicated that Sb and Bs can modulate the microbial ecosystem, and subsequently enhance the health status of broilers.

关键词： saccharomyces boulardii Bacillus subtilis B10 Gastrointestinal tract Microbiota Broiler

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De novo biosynthesis of liquiritin in saccharomyces cerevisiae

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Acta Pharmaceutica Sinica B 2020年第4期10卷 711-721页

作者： Yan Yin Yanpeng Li Dan Jiang Xianan Zhang Wei Gao Chunsheng Liu School of Chinese Pharmacy Beijing University of Chinese MedicineBeijing 102488China School of Traditional Chinese Medicine Capital Medical UniversityBeijing 100069China School of Pharmaceutical Sciences Advanced Innovation Center for Human Brain ProtectionCapital Medical UniversityBeijing 100069China Pharmacy Department Beijing Longfu HospitalBeijing 100010China

Liquiritigenin(LG),isoliquiritigenin(Iso-LG),together with their respective glycoside derivatives liquiritin(LN)and isoliquiritin(Iso-LN),are the main active flavonoids of Glycyrrhiza uralensis,which is arguably the most widely used medicinal plant with enormous demand on the market,including Chinese medicine prescriptions,preparations,health care products and even food.Pharmacological studies have shown that these ingredients have broad medicinal value,including anti-cancer and antiinflammatory effects.Although the biosynthetic pathway of glycyrrhizin,a triterpenoid component from G.uralensis,has been fully analyzed,little attention has been paid to the biosynthesis of the flavonoids of this plant.To obtain the enzyme-coding genes responsible for the biosynthesis of LN,analysis and screening were carried out by combining genome and comparative transcriptome database searches of G.uralensis and homologous genes of known flavonoid biosynthesis pathways.The catalytic functions of candidate genes were determined by in vitro or in vivo characterization.This work characterized the complete biosynthetic pathway of LN and achieved the de novo biosynthesis of liquiritin in saccharomyces cerevisiae using endogenous yeast metabolites as precursors and cofactors for the first time,which provides a possibility for the economical and sustainable production and application of G.uralensis flavonoids through synthetic biology.

关键词： Glycyrrhiza uralensis Liquiritin Isoliquiritin Liquiritigenin Isoliquiritigenin Heterologous synthesis saccharomyces cerevisiae

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Protective effect of saccharomyces boulardii on intestinal mucosal barrier of dextran sodium sulfate-induced colitis in mice

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Chinese Medical Journal 2019年第16期132卷 1951-1958页

作者： Jin-Pei Dong Yue Zheng Ting Wu Qun He Gui-Gen Teng Hua-Hong Wang Department of Gastroenterology Peking University First HospitalBeijing 100034China Department of Urology Peking University First HospitalBeijing 100034China.

Background: The effect and mechanism of saccharomyces boulardii (Sb) in inflammatory bowel disease are unclear. The objective of the study was to evaluate the impact of Sb on intestinal mucosal barrier and intestinal flora in a colitis mouse model. Methods: Forty C57BL/6J male mice were randomly assigned to five groups: normal control group (A), pathologic control group (B), Sb treatment group (C), mesalazine treatment group (D), and Sb combined with mesalazine treatment group (E). Colitis was induced by the addition of 2.5%(wt/vol) dextran sodium sulfate (DSS) in the drinking water ad libitum for 7 days. The general condition, weight change, stool property, and bloody stool level of mice were observed to evaluate the disease activity index. The expression of zona occludens-1 (ZO-1) and occludin in intestinal tissue were measured by immunohistochemistry. The level of tumor necrosis factor-α(TNF-α) and interleukin (IL)-8 in plasma was measured by enzyme linked immunosorbent assay. Inter-cellular tight junctions were observed by transmission electron microscopy. The feces and intestinal contents were collected sterilely, and intestinal flora was analyzed by 16S rRNA sequencing. Results: Compared with group B, Sb reduced the disease activity index and histological score of group C (disease activity index: group B 2.708 ± 0.628, group C 1.542 ± 0.616, PBC = 0.005;histological score: group B 9.875 ± 3.271, group C 4.750 ± 1.832, PBC = 0.005) in DSS-induced colitis in mice. Sb exerted a protect effect on the expression of ZO-1 (group B 2.075 ± 1.176, group C 4.225 ± 1.316, PBC = 0.019) and occludin (group B 2.200 ± 0.968, group C 3.525 ± 1.047, PBC = 0.023). Compared with group B, Sb decreased the level of TNF-α and IL-8 of group C (TNF-α: group B 716.323 ± 44.691 ng/L, group C 521.740 ± 90.121 ng/L, PBC = 0.001;IL-8: group B 128.992 ± 11.475 pg/mL, group C 106.283 ± 15.906 pg/mL, PBC = 0.012). Treatment with Sb preserved the tight junctions and ameliorated microvilli and inter-cellular space. Treatment with Sb also sh

关键词： Inflammatory bowel disease saccharomyces boulardii Intestinal mucosal barrier

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SYNTHESIS AND SECRETION OF HUMAN ATRIAL NATRIURETIC PEPTIDE IN saccharomyces cerevisiae

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Science China Chemistry 1991年第12期34卷 1478-1484页

作者：秦宁魏楠姚锡惠李育阳 Institute of Genetics Fudan University Shanghai 200433 PRC

A chemically synthesized α-hANP gene was inserted into plasmid YFD18, which was an expression-secretion vector of yeast. The recombinant then transformed in the yeast Y33. The expression level of yeast transformants was about 700 μg ANP/L detected by RIA. More than 99% of expression products were secreted in the culture medium. N-terminal analysis of purified product showed that the first 4 amino acid residues of α-hANP were deleted.

关键词： human atrial natriuretic peptide saccharomyces cerevisiae synthesis secretion.

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Increasing isobutanol yield by double-gene deletion of PDC6 and LPD1 in saccharomyces cerevisiae

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Chinese Journal of Chemical Engineering 2016年第8期24卷 1074-1079页

作者： Aili Zhang Yang Li Yuhan Gao Hongxing Jin School of Chemical Engineering and Technology Hebei University of Technology

As a new biofuel, isobutanol has received more attentions in recent years. Because of its high tolerance to higher alcohols, saccharomyces cerevisiae has potential advantages as a platform microbe to produce isobutanol. In this study, we investigated integration effects of enhancing valine biosynthesis by overexpression of ILV2 and BAT2 with eliminating ethanol formation by deletion of PDC6 and decreasing acetyl-Co A biosynthesis by deletion of LPD1 on isobutanol titers. Our results showed that deletion of LPD1 in strains overexpressing BAT2 and ILV2 increased isobutanol titer by 5.3-fold compared with control strain. Additional deletion of PDC6 in lpd1Δ strains carrying overexpressed BAT2 and ILV2 further increased isobutanol titer by 1.5 fold. Overexpression of BAT2 and ILV2 in lpd1Δ strains and pdc6Δ strains decreased ethanol titers. Glycerol titers of the engineered strains did not have greater changes than that of control strain, while their acetic acid titers were higher, perhaps due to the imbalance of cofactors in isobutanol synthesis. Our researches suggest that double-gene deletion of PDC6 and LPD1 in strains overexpressing BAT2 and ILV2 could increase isobutanol production dramatically than single-gene deletion of PDC6 or LPD1. This study reveals the integration effects of overexpression of ILV2/BAT2 and double-gene deletion of LPD1 and PDC6 on isobutanol production, and helps understanding future developments of engineered strains for producing isobutanol.

关键词： saccharomyces cerevisiae Isobutanol Ethanol Pyruvate decarboxylase

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Effects of saccharomyces cerevisiae or boulardii yeasts on acute stress induced intestinal dysmotility

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World Journal of Gastroenterology 2016年第48期22卷 10532-10544页

作者： Christine West Andrew M Stanisz Annette Wong Wolfgang A Kunze McMaster Brain-Body Institute St. Joseph’s Healthcare Department of Biology McM aster University Department of Psychiatry and Behavioural Neurosciences Mc Master University

AIM To investigate the capacity of saccharomyces cerevisiae(S. cerevisiae) and saccharomyces boulardii(S. boulardii) yeasts to reverse or to treat acute stress-related intestinal dysmotility.METHODS Adult Swiss Webster mice were stressed for 1 h in a wire-mesh restraint to induce symptoms of intestinal dysmotility and were subsequently killed by cervical dislocation. Jejunal and colon tissue were excised and placed within a tissue perfusion bath in which S. cerevisiae, S. boulardii, or their supernatants were administered into the lumen. Video recordings of contractility and gut diameter changes were converted to spatiotemporal maps and the velocity, frequency, and amplitude of propagating contractile clusters(PCC) were measured. Motility pre- and post-treatment was compared between stressed animals and unstressed controls. RESULTS S. boulardii and S. cerevisiae helped to mediate the effects of stress on the small and large intestine. Restraint stress reduced jejunal transit velocity(mm/s)from 2.635 ± 0.316 to 1.644 ± 0.238, P < 0.001 and jejunal transit frequency(Hz) from 0.032 ± 0.008 to 0.016 ± 0.005, P < 0.001. Restraint stress increased colonic transit velocity(mm/s) from 0.864 ± 0.183 to 1.432 ± 0.329, P < 0.001 and frequency to a lesser degree. Luminal application of S. boulardii helped to restore jejunal and colonic velocity towards the unstressed controls; 1.833 ± 0.688 to 2.627 ± 0.664, P < 0.001 and 1.516 ± 0.263 to 1.036 ± 0.21, P < 0.001, respectively. S. cerevisiae also had therapeutic effects on the stressed gut, but was most apparent in the jejunum. S. cerevisiae increased PCC velocity in the stressed jejunum from 1.763 ± 0.397 to 2.017 ± 0.48, P = 0.0031 and PCC frequency from 0.016 ± 0.009 to 0.027 ± 0.007, P < 0.001. S. cerevisiae decreased colon PCC velocity from 1.647 ± 0.187 to 1.038 ± 0.222, P < 0.001. Addition of S. boulardii or S. cerevisiae supernatants also helped to restore motility to unstressed values in similar capacity. CONCLUSION There is a potential therapeutic role for S. cerevi

关键词： Intestine saccharomyces cerevisiae saccharomyces boulardii Restraint stress Motility

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Diversity of saccharomyces boulardii CNCM I-745 mechanisms of action against intestinal infections

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World Journal of Gastroenterology 2019年第18期25卷 2188-2203页

作者： Dorota Czerucka Patrick Rampall Department of Human Health Division of Ecosystems and Immunity Center Scientific of Monaco Center Scientific of Monaco

The yeast saccharomyces boulardii CNCM I-745 is one of the probiotics recommended for the prevention of antibiotic-associated diarrhea. Studies conducted in vivo and in vitro demonstrated that in the case of infectious diseases there are two potential sites of action of saccharomyces boulardii CNCM I-745:(1)An action on enteropathogenic microorganisms(adhesion of bacteria and their elimination or an effect on their virulence factors: Toxins, lipopolysaccharide,etc.); and(2) a direct action on the intestinal mucosa(trophic effects, effects on epithelial reconstitution, anti-secretory effects, anti-inflammatory,immunomodulators). Oral administration of saccharomyces boulardii CNCM I-745 to healthy subjects does not alter their microbiota. However, in the case of diseases associated with the use of antibiotics or chronic diarrhea, saccharomyces boulardii CNCM I-745 can restore the intestinal microbiota faster. The interaction of saccharomyces boulardii CNCM I-745 with the innate immune system have been recently demonstrated thus opening up a new therapeutic potential of this yeast in the case of diseases associated with intestinal infections but also other pathologies associated with dysbiosis such as inflammatory diseases.

关键词： saccharomyces boulardii CNCM I-745 Probiotics Yeast Intestinal infection Mechanism

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Engineering the Biosynthesis of Caffeic Acid in saccharomyces cerevisiae with Heterologous Enzyme Combinations

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Engineering 2019年第2期5卷 287-295页

作者： Lanqing Liu Hong Liu Wei Zhang Mingdong Yao Bingzhi Li Duo Liu Yingjin Yuan Key Laboratory of Systems Bioengineering (Ministry of Education) School of Chemical Engineering and Technology Tianjin University Tianjin 300072 China SynBio Research Platform Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) Tianjin University Tianjin 300072 China

Engineering the biosynthesis of plant-derived natural products in microbes presents several challenges, especially when the expression and activation of the plant cytochrome P450 enzyme is required. By recruiting two enzymes—HpaB and HpaC—from several bacteria, we constructed functional 4- hydroxyphenylacetate 3-hydroxylase (4HPA3H) in saccharomyces cerevisiae to take on a role similar to that of the plant-derived cytochrome P450 enzyme and produce caffeic acid. Along with a common tyrosine ammonia lyase (TAL), the different combinations of HpaB and HpaC presented varied capabilities in producing the target product, caffeic acid, from the substrate, L-tyrosine. The highest production of caffeic acid was obtained with the enzyme combination of HpaB from Pseudomonas aeruginosa and HpaC from Salmonella enterica, which yielded up to (289.4 ± 4.6) mg-L1 in shake-flask cultivation. The compatibility of heterologous enzymes within a yeast chassis was effectively improved, as the caffeic acid production was increased by 40 times from the initial yield. Six key amino acid residues around the flavin adenine dinucleotide (FAD) binding domain in HpaB from Pseudomonas aeruginosa were differentiate from those other HpaBs, and might play critical roles in affecting enzyme activity. We have thus established an effective approach to construct a highly efficient yeast system to synthesize non-native hydroxylated phenylpropanoids.

关键词： saccharomyces cerevisiae Caffeic acid Heterologous enzyme Cytochrome P450 Synthetic biology

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Identification of a Long-Chain Fatty Acid Elongase from Nannochloropsis sp. Involved in the Biosynthesis of Fatty Acids by Heterologous Expression in saccharomyces cerevisiae

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Journal of Ocean University of China 2019年第5期18卷 1199-1206页

作者： GUO Minrui CHEN Guogang CHEN Jiluan ZHENG Minggang College of Food Shihezi UniversityShihezi 832000China The First Institute of Oceanography State Oceanic Administration of ChinaQingdao 266061China

The marine microalga Nannochloropsis sp.contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids.A full-length cDNA encoding a long-chain fatty acid elongase,named Ns FAE,was cloned from Nannochloropsis sp..The open reading frame of Ns FAE(GenBank accession no.MF680548)consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 k Da.The deduced polypeptide showed 43%–44%identity to fatty acyl elongases from other algae.RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp.at 72 h(i.e.,during the third growth stage)and the expression was significantly lower in the other four growth stages.Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1p-Ns FAE-CYCt)were transformed into saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system.Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid,linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.

关键词： Nannochloropsis sp. long-chain fatty acid elongase saccharomyces cerevisiae fatty acid production

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