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Design-build-test of recombinant Bacillus subtilis chassis cell by lifespan engineering for robust bioprocesses

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Synthetic and Systems biotechnology 2024年第3期9卷 470-480页

作者： Kexin Ren Qiang Wang Jianghua Chen Hengwei Zhang Zhoule Guo Meijuan Xu Zhiming Rao Xian Zhang key laboratory of industrial biotechnology of the ministry of education School of BiotechnologyJiangnan UniversityWuxiJiangsu214122China Yixing Institute of Food and biotechnology Co. LtdYixing214200China

Microbial cell factories utilize renewable raw materials for industrial chemical production,providing a promising path for sustainable *** subtilis is widely used in industry for its food safety properties,but challenges remain in the limitations of microbial *** study proposes a novel strategy based on lifespan engineering to design robust *** chassis cells to supplement traditional metabolic modification strategies that can alleviate cell autolysis,tolerate toxic substrates,and get a higher mass transfer *** modified chassis cells could produce high levels of L-glutaminase,and tolerate hydroquinone to produceα-arbutin *** a 5 L bioreactor,the L-glutaminase enzyme activity of the final strain CRE15TG was increased to 2817.4±21.7 U m L^(-1),about 1.98-fold compared with that of the wild ***α-arbutin yield of strain CRE15A was increased to 134.7 g L^(-1),about 1.34-fold compared with that of the *** our knowledge,both of the products in this study performed the highest yields reported so *** chassis modification strategy described in this study can Improve the utilization efficiency of chassis cells,mitigate the possible adverse effects caused by excessive metabolic modification of engineered strains,and provide a new idea for the future design of microbial cell factories.

关键词： Bacillus subtilis Chassis cell Lifespan engineering Robustness industrial production

Well-dispersive K2O-KCl alkaline catalyst derived from waste banana peel for biodiesel synthesis

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Green Energy & Environment 2019年第3期4卷 322-327页

作者： Mingming Fan Hao Wu Min Shi Pingbo Zhang Pingping Jiang The key laboratory of Food Colloids and biotechnology Ministry of Education School of Chemical and Material Engineering Jiangnan University

In this work, a novel alkaline catalyst was synthesized by an economical and effective method of roasting waste banana peel. From XRD, FTIR, SEM, EDS, TGA and CO2-TPD characterization, it was proved that the calcined banana peel catalyst showed the strong alkalinity and well dispersity in microstructure, and K2O-KCl as the main active contents. The calcined banana peel catalyst showed better catalytic performance than the catalysts by physical mixing of K2O and KCl, which was due to good dispersibility of K2O-KCl formed during decomposing of carbon fiber by calcination of banana peel. Furthermore, the calcined banana peel catalyst also performed well in both water-resistant ability and recyclability, indicating their potential for biodiesel production from an efficient, robust, and low-cost catalyst.

关键词： Biodiesel Banana peel Basic catalyst Transesterification

Preparation and Crystal Structure of 12β,15α-dihydroxyprogesterone

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Chinese Journal of Structural Chemistry 2011年第12期30卷 1687-1690页

作者：孙华沈妍君赵旭王敏 key laboratory of industrial Microbiology Ministry of EducationCollege of BiotechnologyTianjin University of Science and Technology

Progesterone（1） was biotransformed into the title compound 12β,15α-dihydroxy-progesterone（2） with Colletotrichum lini *** biotransformation process was monitored with *** structure of 2 was determined by 1H NMR,13C NMR,ESI-MS and single-crystal X-ray *** crystal of the compound belongs to orthorhombic,space group P212121 with a = 8.0671（9）,b = 12.3970（15）,c = 18.532（3） ,Z = 4,V = 1853.3（4）3,Dc = 1.242 mg/m3,μ = 0.084 mm-1,F（000） = 752,R = 0.0373 and wR = ***-crystal X-ray diffraction analysis reveals that a 1D supramolecular structure of 2 has been constructed by multiply intermolecular O（2）-H（2）…O（4） and O（3）-H（3）…O（2） H-bonding interactions.

关键词： crystal structure progesterone 12β 15α-dihydroxyprogesterone biotransformation

Preparation and Crystal Structure of 15α-Hydroxyl-oleanolic Acid

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Chinese Journal of Structural Chemistry 2010年第12期29卷 1798-1801页

作者：孙华刘婷沈妍君张黎明王敏 key laboratory of industrial Microbiology Ministry of EducationCollege of BiotechnologyTianjin University of Science and Technology

The compound 15α-hydroxyl-oleanolic acid was biotransformed from oleanolic acid with Colletotrichum lini AS3.4486,and its structure was characterized by 1H NMR,13C NMR,HR-ESI-MS and single-crystal X-ray *** crystal of the title compound belongs to monoclinic,space group R3 with a=34.255（5）,b=34.255（5）,c=6.8671（14）,β=90°,Z=9,V=6978（2）3,Mr=490.70,Dc=1.051 mg/m3,μ=0.069 mm-1,F（000）=2430,R=0.0375 and wR=*** title compound is stacked into a three-dimensional network through hydrogen bonds.

关键词： crystal structure oleanolic acid 15α-hydroxyl-oleanolic acid biotransformation

Whole‑cell catalytic synthesis of trehalose by Corynebacterium glutamicum displaying trehalose synthase on its cell surface

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Systems Microbiology and Biomanufacturing 2023年第3期3卷 489-497页

作者： Ming Fang Qiang Wang Zhi Wang Ruiqi Lu Rufan Xing Xian Zhang Zhiming Rao key laboratory of industrial biotechnology of ministry of education School of BiotechnologyJiangnan UniversityWuxi 214122Jiangsu ProvincePeople’s Republic of China

Trehalose is a disaccharide with many applications in cosmetics,refrigeration,and *** synthase is a significant enzyme in trehalose *** coli is usually used to express this enzyme *** *** is a pathogenic strain,we chose Corynebacterium glutamicum ATCC13032 as an engineering strain in this study for food safety *** of its poor permeability,we constructed two recombinant *** strains using two anchor proteins,PorH,and short-length NCgl1337,to anchor trehalose synthase from Streptomyces coelicolor on the cell surface and synthesize trehalose directly from *** on enzymatic properties indicated that NCgl1337S–ScTreSK246A had better enzyme activity and thermal stability than the free *** optimizing the whole-cell transformation,the optimal transformation condition was 35°C,pH 7.0,and OD600 of *** this condition,the conversion rate of 300 g/L maltose reached 69.5%in a 5 L *** relative conversion rate was still above 75%after repeated five times.

关键词： Trehalose synthase Corynebacterium glutamicum Surface display Whole-cell catalyses

Influence of aeration intensity on the performance of A/0-type sequencing batch MBR system treating azo dye wastewater

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Frontiers of Environmental Science & Engineering 2011年第4期5卷 615-622页

作者： Xinhua WANG Jingmei LI Xiufen LI Guocheng DU laboratory of Environmental biotechnology School of Environmental and Civil EngineeringJiangnan UniversityWuxi 214122China key laboratory of industrial biotechnology Ministry of EducationJiangnan UniversityWuxi 214122China

Among the numerous parameters affecting the membrane bioreactor(MBR)performance,the aeration intensity is one of the most important *** the present investigation,an anoxic/aerobic-type(A/O-type)sequencing batch MBR system,added anoxic process as a pretreatment to improve the biodegradability of azo dye wastewater,was investigated under different aeration intensities and the impact of the aeration intensity on effluent quantity,sludge properties,extracellular polymeric substances(EPS)amount generated as well as the change of permeation flux were *** lower nor higher aeration intensities could improve A/O-type sequencing batch MBR *** results showed 0.15 m^(3)·h^(-1)aeration intensity was promising for treatment of azo dye wastewater under the conditions *** this aeration intensity,chemical oxygen demand(COD),ammonium nitrogen and color removal as well as membrane flux amounted to 97.8%,96.5%,98.7%and 6.21 L·m^(-2)·h^(-1),*** effluent quality,with 25.0 mg·L^(-1)COD,0.84 mg·L^(-1)ammonium nitrogen and 8 chroma,could directly meet the reuse standard in *** the meantime,the sludge relative hydrophobicity,the bound EPS,soluble EPS and EPS amounts contained in the membrane fouling layer were 70.3%,52.0 mg·g^(-1)VSS,38.8 mg·g^(-1)VSS and 90.8 mg·g^(-1)VSS,respectively,which showed close relationships to both pollutant removals and membrane flux.

关键词： batch membrane bioreactor azo dye aeration intensity extracellular polymeric substances sludge properties

Effect of Lanthanum (Ⅲ) on Reactive Oxygen Metabolism of Soybean Seedlings under Supplemental UV-B Irradiation

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Journal of Rare Earths 2007年第3期25卷 352-358页

作者：闫生荣黄晓华周青 key laboratory of industrial biotechnology Ministry of EducationSouthern Yangtze University College of Chemistry and Environment Science Nanjing Normal University

The effect of lanthanum （Ⅲ） on reactive oxygen metabolism of soybean seedlings under elevated ultraviolet-B radiation（UV-B：280~320 nm）at 0.15 and 0.45 W·cm-2 levels respectively was studied through hydroponics in the *** permeability and contents of malonadialdehyde（MDA）,hydrogen peroxide（H2O2）,and proline gradually increased during the imposition of UV-B radiation and subsequently decreased during recovery from UV-B *** dynamic tendency of catalase（CAT）activity was similar to that of the above four *** activity of peroxidase（POD）initially increased,then remained at a high level,and finally dropped steeply when soybean seedlings were exposed to a low dosage of UV-B ***,POD activity rose throughout and declined slightly on the eleventh day when soybean seedlings were stressed by a high *** the addition of La （Ⅲ） of 20 mg·L-1,the rising tendency of plasmolemma permeability and contents of MDA,H2O2,and proline were slowed down during the stress period,whereas the declining speed was accelerated during the recovery *** activities of CAT and POD were higher than those without La （Ⅲ） in all *** suggested that the regulative effect of La （Ⅲ） on antioxidant enzymes such as CAT and POD could strengthen their capacities to scavenge reactive oxygen species（ROS）,decrease contents of MDA and proline,and maintain normal plasmolemma *** more,the protective potential of La （Ⅲ） was better under low UV-B radiation than under a high one.

关键词： lanthanum (Ⅲ) ultraviolet-B radiation soybean seedlings metabolism of reactive oxygen species CATactivity POD activity

Synthesis and immunological study of a glycosylated wall teichoic acid-based vaccine against Staphylococcus aureus

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Chinese Chemical Letters 2023年第4期34卷 193-197页

作者： Peng Shen Han Lin Yikai Bao Haofei Hong Zhimeng Wu key laboratory of Carbohydrate Chemistry&biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi 214122China

Staphylococcus aureus wall teichoic acids(WTAs) are attractive targets for antibacterial vaccine development. In this study, three core glycosylated WTA structure, including α-1,4-Glc NAc, β-1,4-Glc NAc andβ-1,3-Glc NAc modified ribitol phosphates containing a linker are chemically synthesized and conjugated with tetanus toxin(TT) carrier protein as vaccine candidates. In vivo immunological studies demonstrate that the synthesized glycosylated WTAs display high immunogenicity and all conjugates provoke strong immune responses and elicit high levels of specific IgG antibodies against the Glc NAc-modified WTA. Furthermore, antibodies elicited by the vaccine candidates remain the capability to recognize S. aureus cells and display significant opsonophagocytic activity to clear S. aureus. This study demonstrates that the core structure of glycosylated WTAs are effective antigens for constructing anti-S. aureus vaccines to prevent and control S. aureus infections.

关键词： WTAs Staphylococcus aureus GlcNAc modifications Glycoconjugates Antibacterial vaccine

Synthesis of DNP-modified GM3-based anticancer vaccine and evaluation of its immunological activities for cancer immunotherapy

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Chinese Chemical Letters 2021年第12期32卷 4041-4044页

作者： Han Lin Haofei Hong Lipeng Feng Jie Shi Zhifang Zhou Zhimeng Wu key laboratory of Carbohydrate Chemistry&biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi 214122China

Tumor-associated carbohydrate antigens(TACAs) are attractive targets for vaccine development. In this context, we described a strategy combining artificial TACA and glycoengineering for cancer vaccine development. A 2,4-ditrophenyl(DNP)-modified GM3 intermediate was synthesized chemoenzymatically and conjugated to keyhole limpet hemocyanin(KLH), and the resulting bioconjugate was tested for its potential as a vaccine candidate. Mice immunological studies revealed that the DNP-modified GM3(GM3-NHDNP) analog elicited strong and rapid immune responses by recruiting anti-DNP antibodies to facilitate the targeted delivery of the vaccine construct to antigen processing cells(APCs). Moreover, the endogenously produced anti-DNP antibodies, together with the elicited antibodies against GM3-NHDNP, may synergistically promote tumor binding and cancer cell death when the cancer cell surfaces are glycoengineered to express the GM3-NHDNP antigen.

关键词： GM3 Cancer vaccine Anti-DNP antibodies Glycoengineering Tumor-associated carbohydrate antigen