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De novo biosynthesis of 8‑prenylnaringenin in Saccharomyces cerevisiae improved by screening and engineering of prenyltransferases and precursor pathway

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Systems Microbiology and Biomanufacturing 2023年第4期3卷 647-658页

作者： Chaojie Guo Yongkun Lv Hongbiao Li Jingwen Zhou Sha Xu National Engineering laboratory for Cereal Fermentation Technology(NELCF) Jiangnan University1800 Lihu RoadWuxi 214122JiangsuChina Key laboratory of Industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan University1800 Lihu RoadWuxi 214122JiangsuChina Science Center for Future Foods Jiangnan University1800 Lihu RoadWuxi 214122JiangsuChina School of Chemical Engineering Zhengzhou UniversityZhengzhou 450001HenanChina Jiangsu Provisional Research Center for bioactive Product Processing Technology Jiangnan University1800 Lihu RoadWuxi 214122JiangsuChina

8-Prenylnaringenin(8-PN)is a valuable medical phytoestrogen,which is a precursor to many prenylated ***-ever,the availability of 8-PN is limited by inefficient prenyltransferases(PTs)and inadequate substrate precursor levels in microbial ***,six PTs from different sources and their truncated cognates were expressed in a(2S)-naringenin producing *** SfN8DT-1 derived from Sophora flavescens and its truncated cognate,tSfN8DT-1,could synthe-size ***,tSfN8DT-1 was engineered by multiple sequence alignment and a mutant tSfN8DT-1^(Q12E)with greater catalytic activity was ***,key genes,tHMGR and IDI1,of the mevalonate(MVA)pathway were overexpressed using a copy number combinatorial strategy,which greatly improved 8-PN titer by 368.75%.Fourth,a predicted structure of tSfN8DT-1^(Q12E)was used for molecular docking and virtual saturation *** key residues,P229 and N305,were identified and saturation mutagenesis on these two sites resulted in an improved mutant *** best-performing mutant,tSfN8DT-1^(Q12EN305M),produced 49.35±0.05 mg/L(5.57±0.01 mg/g DCW)8-PN in a shaking ***,101.40±2.55 mg/L of 8-PN was obtained in a 5-L bioreactor,which is the greatest titer reported to date for *** study combined metabolic engineering and protein engineering methods to enhance precursor supplements and improve the catalytic ability of *** production of 8-PN in Saccharomyces cerevisiae was greatly increased through these methods,which may provide a feasible strategy for the biosynthesis of prenylated flavonoids.

关键词： Saccharomyces cerevisiae Prenyltransferases Prenylated flavonoid Protein engineering 8-Prenylnaringenin

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Distinct co‑occurrence patterns and driving forces of abundant and rare bacterial communities in the multispecies solid‑state fermentation process of cereal vinegar

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Systems Microbiology and Biomanufacturing 2022年第2期2卷 317-330页

作者： Ming‑Ye Peng Zhen‑Ming Lu Xiao‑Juan Zhang Ting Huang Yong‑Jian Deng Li‑Juan Chai Jin‑Song Shi Zheng-Hong Xu Key laboratory of Industrial biotechnology of Ministry of Education School of BiotechnologyJiangnan UniversityWuxi 214122People’s Republic of China National Engineering laboratory for Cereal Fermentation Technology Jiangnan University1800 Lihu AvenueWuxi 214122People’s Republic of China School of Pharmaceutical Science Jiangnan UniversityWuxi 214122People’s Republic of China Jiangsu Engineering Research Center for bioactive Products Processing Technology Jiangnan UniversityWuxi 214122People’s Republic of China National Engineering Research Center of Solid-State Brewing Luzhou 646000People’s Republic of China

Multispecies solid-state fermentation is a traditional processing technique for the traditional Chinese food,such as cereal vinegar,Baijiu,***,few abundant and many rare microbes were involved in such processes,and the necessity and roles of the latter are less *** the co-occurrence patterns of abundant and rare bacterial community and abiotic factors infuencing their community assembly were investigated in acetic acid fermentation following starter inoculation,using Zhenjiang aromatic vinegar as a model *** taxa that contribute to the function of accumulating acid exhibited a ubiquitous distribution while the distribution of rare taxa along the fermentation process *** species composition of the rare taxa signifcantly altered,but abundant taxa were maintained after ***,the diversity of rare taxa changed more signifcantly than that of abundant *** abundant and rare sub-communities,which were contributed more with species turnover than species richness,were demonstrated to be driven by pH,acetic acid,ammonium nitrogen,and *** processes regulated the assembly of both sub-communities,but more prominent on rare ***-occurrence network was more governed by rare sub-communities,and the co-variations between microbial communities were predominantly positive,implying that rare taxa played more important role in the fermentation stability and network ***,seven network connectors were identifed,and three of them belonged to rare *** microbes of diferent modules were enriched at particular phases of *** results demonstrate the ecological signifcance of rare bacteria and provide new insights into understanding the abiotic factors infuence microbial structure in traditional fermented foods.

关键词： Solid-state fermentation Rare sub-community Community assembly Co-occurrence network Microbial interactions

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Development of a novel platform for recombinant protein production in Corynebacterium glutamicum on ethanol

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Synthetic and Systems biotechnology 2022年第2期7卷 765-774页

作者： Xinyu Yu Xiuxia Liu Xiong Gao Xunxun Luo Yankun Yang Ye Li Chunli Liu Chong Zhang Zhonghu Bai Key laboratory of Industrial biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxiChina National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing Jiangnan UniversityWuxiChina Division of Life Science The Hong Kong University of Science and TechnologyClear Water BayHong KongChina Jiangsu Provincial Research Center for bioactive Product Processing Technology Jiangnan UniversityWuxiChina MOE Key laboratory for Industrial Biocatalysis Institute of Biochemical EngineeringDepartment of Chemical EngineeringTsinghua UniversityBeijingChina Center for Synthetic and Systems Biology Tsinghua UniversityBeijingChina

Corynebacterium glutamicum represents an emerging recombinant protein expression factory due to its ideal features for protein secretion,but its applicability is harmed by the lack of an autoinduction system with tight regulation and high ***,we propose a new recombinant protein manufacturing platform that leverages ethanol as both a delayed carbon source and an ***,we reanalysed the native inducible promoter PICL from the acetate uptake operon and found that its limited capacity is the result of the inadequate translation initial *** two strategies of bicistronic design and ribozyme-based insulator can ensure the high activity of this ***,through transcriptional engineering that alters transcription factor binding sites(TFBSs)and the first transcribed sequence,the truncated promoter PA256 with a dramatically higher transcription level was *** producing the superfolder green fluorescent protein(sfGFP)under 1%ethanol conditions,PA256 exhibited substantially lower protein accumulation in prophase but an approximately 2.5-fold greater final yield than the strong promoter *** superior expression mode was further validated using two secreted proteins,camelid antibody fragment(VHH)and endoxylanase(XynA).Furthermore,utilizing CRISPRi technology,ethanol utilization blocking strains were created,and PA256 was shown to be impaired in the phosphotransacetylase(PTA)knockdown strains,indicating that ethanol metabolism into the tricarboxylic acid cycle is required for PA256 ***,this platform was applied to produce the“de novo design”protein NEO-2/15,and by introducing the N-propeptide of CspB,NEO-2/15 was effectively secreted with the accumulation 281 mg/L obtained after 24 h of shake-flask *** the best of our knowledge,this is the first report of NEO-2/15 secretory overexpression.

关键词： Corynebacterium glutamicum Ethanol Transcriptional engineering Secretory protein overexpression NEO-2/15

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Isolation,identification and characterization of an algicidal bacterium from Lake Taihu and preliminary studies on its algicidal compounds

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Journal of Environmental Sciences 2012年第10期24卷 1823-1831页

作者： Chuan Tian Xianglong Liu Jing Tan Shengqin Lin Daotang Li Hong Yang State Key laboratory of Microbial metabolism School of Life Science and BiotechnologyShanghai Jiao Tong University800 Dongchuan Rd.Shanghai 200240China

In an effort to identify a bio-agent capable of controlling cyanobacterial blooms, we isolated a bacterial strain, A27, which exhibited strong algicidal activity against the dominant bloom-forming species of Microcystis aeruginosa in Lake Taihu. Based on 16S rRNA gene sequence analysis, this strain belongs to the genus Exiguobacterium. This is the first report of an algicidal bacterial strain belonging to the genus Exiguobacterium. Strain A27 exhibited algicidal activity against a broad range of cyanobacteria, but elicited little or no algicidal activity against the two green algal strains tested. The algicidal activity of strain A27 was shown to be dependent on the density of the bacteria and to have a threshold density of 1.5×106 CFU/mL. Our data also showed that the algicidal activity of strain A27 depended on different growth stages ofMicrocystis aeruginosa （exponential ≈ lag phase 〉 early stationary） rather than that of the bacterium itself. Our results also suggested the algicidal activity of strain A27 occurred via the production of extracellular algicidal compounds. Investigation of the algicidal compounds revealed that there were at least two different algicidal compounds produced by strain A27. These results indicated that strain A27 has great potential for use in the control of outbreaks of cyanobacterial blooms in Lake Taihu.

关键词： algicidal bacteria freshwater cyanobacteriat bloom Microcystis aeruginosa Exiguobacterium sp. A27

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The cell cycle inhibitor P21 promotes the development of pulmonary fibrosis by suppressing lung alveolar regeneration

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Acta Pharmaceutica Sinica B 2022年第2期12卷 735-746页

作者： Xiaoxi Lv Chang Liu Shanshan Liu Yunxuan Li Wanyu Wang Ke Li Fang Hua Bing Cui Xiaowei Zhang Jiaojiao Yu Jinmei Yu ZhuoWei Hu Immunology and Cancer Pharmacology Group State Key Laboratory of Bioactive Substance and Function of Natural MedicinesInstitute of Materia MedicaChinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China Drug Clinical Trial Institution Children’s HospitalCapital Institute of PediatricsBeijing 100020China Institute of Medicinal biotechnology Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100050China

The cell cycle inhibitor P21 has been implicated in cell senescence and plays an important role in the injury-repair process following lung *** fibrosis(PF)is a fibrotic lung disorder characterized by cell senescence in lung alveolar epithelial *** this study,we report that P21 expression was increased in alveolar epithelial type 2 cells(AEC2 s)in a time-dependent manner following multiple bleomycin-induced *** injury of AEC2 s resulted in telomere shortening and triggered P21-dependent cell ***2 s with elevated expression of P21 lost their self-renewal and differentiation *** particular,elevated P21 not only induced cell cycle arrest in AEC2 s but also bound to P300 andβ-catenin and inhibited AEC2 differentiation by disturbing the P300-β-catenin ***,senescent AEC2 s triggered myofibroblast activation by releasing profibrotic *** of P21 restored AEC2-mediated lung alveolar regeneration in mice with chronic *** results of our study reveal a mechanism of P21-mediated lung regeneration failure during PF development,which suggests a potential strategy for the treatment of fibrotic lung diseases.

关键词： P21 Cell senescence Alveolar epithelial type 2 cells Pulmonary fibrosis Alveolar regeneration Bleomycin Cell cycle arrest P300-β-catenin complex

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Optimizing production ofasperolide A,a potential anti-tumor tetranorditerpenoid originally produced by the algal-derived endophytic fungus Aspergillus wentii EN-48

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Chinese Journal of Oceanology and Limnology 2017年第3期35卷 658-663页

作者：徐蕊李晓明徐刚明王斌贵 laboratory of Marine Biology and biotechnology Qingdao National Laboratory for Marine Science and TechnologyKey Laboratory of Experimental Marine BiologyInstitute of OceanologyChinese Academy of SciencesQingdao 266071China University of Chinese Academy of Sciences Beijing 100049China

The marine algal-derived endophytic fungus Aspergillus wentii EN-48 produces the potential anti-tumor agent asperolide A, a tetranorlabdane diterpenoid active against lung cancer. However, the fermentation yield of asperolide A was very low and only produced in static cultures. Static fermentation conditions of A. wentii EN-48 were optimized employing response surface methodology to enhance the production of asperolide A. The optimized conditions resulted in a 13.9-fold yield enhancement, which matched the predicted value, and the optimized conditions were successfully used in scale-up fermentation for the production of asperolide A. Exogenous addition of plant hormones （especially 10 pmol/L methyl jasmonate） stimulated asperolide A production. To our knowledge, this is first optimized production of an asperolide by a marine-derived fungus. The optimization is effective and valuable to supply material for further anti-tumor mechanism studies and preclinical evaluation of asperolide A and other norditerpenoids.

关键词： Aspergillus wentii asperolide A anti-tumor agent fermentation optimization

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Characterization of aroma profiles and aroma-active compounds in high-salt and low-salt shrimp paste by molecular sensory science

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Food Bioscience 2022年第1期45卷 174-184页

作者： Jing Yu Kuan Lu Jiwei Zi Xihong Yang Wancui Xie College of Marine Science and Biological Engineering Qingdao University of Science and TechnologyQingdaoShandong266042PR China Qingdao Engineering Research Center of Marine bioactive Substances and Precision Nutrition QingdaoShandong266042PR China Shandong Provincial Key laboratory of Biochemical Engineering Shandong266042PR China Qingdao Keda Future biotechnology Co. Ltd.QingdaoShandong266042PR China

Flavor characteristics of raw shrimp,high-salt and low-salt shrimp paste were evaluated and compared by sensory analysis,headspace-gas chromatography-ion mobility spectrometry(HS-GC-IMS)and solid-phase microextraction-gas chromatography-mass spectrometry(SPME-GC-MS).In addition,aroma profiles and aroma-active compounds in these three samples were analyzed by solvent-assisted flavor evaporation-gas chromatography-olfactometry-mass spectrometry(SAFE-GC-O-MS).Results of sensory evaluation revealed that these three samples showed significant differences in sensitivity to the W2W,W2S,W1S and W1W sensors of the electronic nose(pcompounds,short chain alkanes,and sulfides,*** of sensory parameters indicated tha high-salt shrimp paste had a strong roasted and meaty aroma,while low-salt shrimp paste showed an umami aroma more similar to raw shrimp.A total of 118 volatile compounds were identified using the three afore-mentioned gas chromatography *** them,37 aroma-active compounds were detected by ***-salt shrimp was shown to contain more acids,aldehydes and heterocyclic compounds,which resembled cheese,roasted and meaty *** contrast,low-salt shrimp paste contained more esters,which was similar to the umami aroma profile of raw ***,pyrazines were only produced in fermented shrimp paste samples,suggesting that they were produced during the fermentation process and they may contribute unique roasted and meaty aromas,which were important for the overall aroma characteristics of shrimp paste.

关键词： shrimp Paste Salt content Molecular sensory science Aroma-active compounds

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Phylogenetic diversity and specificity of bacteria associated with Microcystis aeruginosa and other cyanobacteria

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Journal of Environmental Sciences 2009年第11期21卷 1581-1590页

作者： SHI Limei CAI Yuanfeng YANG Hualin XING Peng LI Pengfu KONG Lingdong KONG Fanxiang State Key laboratory of Pharmaceutical biotechnology School of Life Sciences Nanjing University Nanjing 210093 China State Key laboratory of Lake Science and Environment Nanfing Institute of Geography and Limnology Chinese Academy of Sciences Nanfing 210008 China

Interactions between bacteria and cyanobacteria have been suggested to have a potential to influence harmful algal bloom dynamics; however, little information on these interactions has been reported. In this study, the bacterial communities associated with five strains of Microcystis aeruginosa, three species of other Microcystis spp., and four representative species of non-Microcystis cyanobacteria were compared. Bacterial 16S rDNA fragments were amplified and separated by denaturing gradient gel electrophoresis （DGGE） followed by DNA sequence analysis. The similarities among bacterial communities associated with these cyanobacteria were compared to the digitized DGGE profiles using the cluster analyses. The bacterial community structure of all cyanobacterial cultures differed. Cluster analysis showed that the similarity values among M. aeruginosa cultures were higher than those of other cyanobacterial cultures. Sequence analysis of DGGE fragments indicated the presence of bacteria including, Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacteroidetes and Actinobacteria in the cyanobacterial cultures. Members of the Sphingomonadales were the prevalent group among the Microcystis-associated bacteria. The results provided further evidence for species-specific associations between cyanoabcteria and heterotrophic bacteria, which are useful for understanding interactions between Microcystis and their associated bacteria.

关键词： bacterial diversity specificity Microcystis denaturing gradient gel electrophoresis

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Structural Basis for Complementary and Alternative Medicine:Phytochemical Interaction with Non-Structural Protein 2 Protease-A Reverse Engineering Strategy

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Chinese Journal of Integrative Medicine 2015年第6期21卷 445-452页

作者： G.Koushik Kumar G.Prasanna T.Marimuthu N.T.Saraswathi Molecular Biophysics laboratory School of Chemical and BiotechnologyShanmuga Arts Science Technology Research Academy University Assistant Medical Officer Goverment Hospital

Objective： To understand the druggability of the bioactive compounds from traditional herbal formulations ＂Nilavembu Kudineer＂ and ＂Swasthya Raksha Amruta Peya＂ to heal chikungunya virus （CHIKV） infection. Methods： The efficiency of twenty novel chemical entities from ＂Nilavembu Kudineer＂ and ＂Swasthya Raksha Amruta Peya＂ to inhibit CHIKV infection in silico were evaluated. Ligands were prepared using Ligprep module of Schr0dinger. Active site was identified using SiteMap program. Grid box was generated using receptor grid generation wizard. Molecular docking was carried out using Grid Based Ligand Docking with Energetics （GLIDE） program. Results： Molecular docking studies showed that among twenty compounds, andrographoside, deoxyandrographoside, neoandrographolide, 14-deoxy-11-oxoandrographolide, butoxone and oleanolic acid showed GLIDE extra precision （XP） score of-9.10,-8.72, -8.25,-7.38,-7.28 and -7.01, respectively which were greater than or comparable with chloroquine （reference compound） XP score （-7.08） and were found to interact with the key residues GLLI 1043, LYS 1045, GLY 1176, LEU 1203, HIS 1222 and LYS 1239 which were characteristic functional unit crucial for replication of CHIKV. Conclusion： The binding affinity and the binding mode of chemical entities taken from herbal formulations with non-structural protein 2 protease were understood and our study provided a novel strategy in the development and design of drugs for CHIKV infection.

关键词： chikungunya virus non-structural protein 2 protease Grid Based Ligand Docking with Energetics score Nilavembu Kudineer Swasthya Raksha Amruta Peya

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The Impact of Inoculum Preparation Media on Pollutant Removal through Phycoremediation of Agricultural Drainage Water by Desmodesmus sp.

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国际实验植物学杂志(英文) 2023年第10期92卷 2875-2890页

作者： Asmaa Salah Hoda Sany Abo El-Khair B.El-Sayed Reham M.El-Bahbohy Heba I.Mohamed Ayman Amin Plant Physiology Division Department of Agricultural BotanyFaculty of AgricultureCairo UniversityGizaP.O.Box 12613Egypt algal biotechnology Unit National Research CentreDokkiGiza12622Egypt Biological and Geological Science Department Faculty of EducationAin Shams UniversityCairo11341Egypt

Water is the most essential natural resource for the future *** production is extensively water-dependent and a significant polluter of water ***,this work investigated the effect of two different preparation media[Bold’s Basal Medium(BBM)and Domiati cheese whey(DCW)]for agricultural drainage water(ADW)*** treatments were incubated for 6 *** to the results of biomass productivity,specific growth rate,photosynthetic pigments,and biochemical composition,Desmodesmus *** grow in drainage water without *** two treatments significantly reduced the concentration of nitrate,phosphate,chemical oxygen demand,and sodium in ***,using cheese whey with BBM as inoculum preparation media enhanced the lipid accumulation in the algal biomass(25.4%w/w)and gave the best biodiesel properties among ***,combining remediation of drainage water with microalgae mass production can achieve the sustainability of agriculture.

关键词： Desmodesmus sp. drainage water nutrients recycle photosynthetic pigments phycoremediation water scarcity

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