Article
Biochemistry & Molecular Biology
Ziqiang Gu, Chen Ning, Zhemin Liu, Qingping Liang, Xiaodan Fu, Ming Tian, Changliang Zhu, Haijin Mou
Summary: This study successfully expressed nattokinase in Bacillus subtilis and improved its enzyme activity by optimizing the start codon and screening signal peptides. Furthermore, constructing a dual-promoter expression vector and optimizing fermentation conditions further enhanced the expression level of nattokinase. This lays the foundation for the industrial development of nattokinase as a thrombolytic reagent.
PROCESS BIOCHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Yuan Yuan, Demin Kong, Jing Wu, Lingqia Su
Summary: In this study, the optimal promoter SpoVG and signal peptide YqxI were obtained and the chitinase BlChiA from Bacillus licheniformis was successfully recombinantly expressed in Bacillus subtilis. Mutant libraries were constructed and mutants with enhanced activities were obtained. The mutant E336K/A472T exhibited the highest expression level of BlChiA in Bacillus subtilis and also showed greater acid adaptability. These findings provide a foundation for further exploration of the enzyme's function and industrial application.
PROCESS BIOCHEMISTRY
(2023)
Article
Agriculture, Multidisciplinary
Guangwen Zhang, Min Lin, Miaomiao Qin, Qianmei Xie, Mingzhi Liang, Jinfei Jiang, Huilin Dai, Siqi Xu, Saixiang Feng, Ming Liao
Summary: Bacillus subtilis was identified as a suitable host for the production of MccJ25 and MccY. High-level production was achieved through promoter optimization, host strain selection, and recombinant expression. This study provides engineered strains of B. subtilis for antibacterial therapy and food preservation, which are free of antibiotic resistance markers, inducer-free, sporulation-deficient, and without the negative effects of endotoxins.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Jing Lu, Yu Zhao, Yu Cheng, Rong Hu, Yaowei Fang, MingSheng Lyu, Shujun Wang, Zhaoxin Lu
Summary: This study enhanced the expression of acetaldehyde dehydrogenase from Issatchenkia terricola in Bacillus subtilis by optimizing signal peptides, promoters, and growth conditions. The optimized signal peptide was yqzG and the combination of P43-Pglv promoters showed the best performance. Through one-factor-at-a-time optimization, the highest production of istALDH was achieved.
Article
Microbiology
Laichuang Han, Qiaoqing Chen, Jie Luo, Wenjing Cui, Zhemin Zhou
Summary: In this study, a Glycerol Inducible Expression System (GIES) was constructed in B. subtilis using the antiterminator protein GlpP and the GlpP-regulated promoter P-glpD. The GIES was able to express genes in an autoinducible manner based on the sequential utilization of glucose and glycerol. This system showed high efficiency and potential for large-scale protein production. The use of the antiterminator protein as a regulator in synthetic biology was also demonstrated.
MICROBIOLOGY SPECTRUM
(2022)
Article
Biotechnology & Applied Microbiology
Dongbang Yao, Xudong Han, Huanhuan Gao, Bin Wang, Zemin Fang, He Li, Wei Fang, Yazhong Xiao
Summary: In this study, the extracellular production level of Pontibacillus sp. ZY raw starch-degrading a-amylase (AmyZ1) in B. subtilis was enhanced by expression regulatory element modification and fermentation optimization. The resulting recombinant strain WBZ-VY-B-R1 showed the highest reported production level for recombinant RSDA production.
MICROBIAL CELL FACTORIES
(2023)
Article
Agriculture, Multidisciplinary
Huabiao Miao, Yuanyuan Zhe, Xia Xiang, Yan Cao, Nanyu Han, Qian Wu, Zunxi Huang
Summary: Protease EA1 is a Ca2+- and Mg2+-dependent hyperthermostable neutral protease with excellent heat resistance, and its production can be improved through mutagenesis. It has potential applications in high-temperature industries.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Biotechnology & Applied Microbiology
Zidong Jiang, Lujia Zhang, Wei Zhou, Hui Li, Yuanyi Li, Weichuang Qin, Fengqing Wang, Dongzhi Wei, Bei Gao
Summary: In this study, a binding structure between SecA and a signal peptide was established, and a grinding approach was used to modify them, leading to improved protein secretion efficiency. A bidirectional grinding strategy was also developed to enhance the secretion system, resulting in a significantly increased protein activity. This research provides a novel and promising strategy for optimizing protein secretion and production in B. subtilis.
BIOCHEMICAL ENGINEERING JOURNAL
(2023)
Article
Multidisciplinary Sciences
Sarah Gingichashvili, Doron Steinberg, Ronit Vogt Sionov, Osnat Feuerstein, Noa E. Cohen
Summary: In this study, computational analytics were used to quantify the morphological response of Bacillus subtilis to the proximity of an antimicrobial source. The results revealed that the inner core, characterized by wrinkled formations, is more preserved compared to the periphery. Additionally, a crescent-shaped colony morphology and changes in the growth substrate of macrocolonies exposed to the antimicrobial were observed.
SCIENTIFIC REPORTS
(2022)
Article
Biochemistry & Molecular Biology
Seunghyun Sim, Yue Hui, David A. Tirrell
Summary: The incorporation of living cells into materials presents both challenges and possibilities. In this study, a new class of 3D-printable materials based on poly(N-hydroxymethylacrylamide) is developed, which allows the cross-linking of spore-forming bacteria into a polymeric scaffold. These embedded cells and spores can become metabolically active and produce proteins after re-swelling in nutrient-rich media. The leak-free scaffold enables protein production while preventing cell escape. This research demonstrates the potential utility of living composite materials in various applications.
Article
Biochemistry & Molecular Biology
Sarika Agrawal, Uttam Kumar Jana, Naveen Kango
Summary: The study cloned and overexpressed the L-asparaginase from Bacillus subtilis ETMC-2, demonstrating its high activity and stability. Physico-chemical properties predictions suggest potential applications for this enzyme.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Biochemistry & Molecular Biology
Kang Zhang, Lingqia Su, Jing Wu
Summary: Bacillus subtilis is a popular host for recombinant protein production due to its many attributes, but residual proteases can limit protein yield. This study successfully increased the production of industrial enzyme pullulanase by enhancing extracellular folding and optimizing its signal peptide. Disrupting the dltB gene of B. subtilis WS9 increased pullulanase production by 49% and identifying a more effective signal peptide further increased extracellular pullulanase activity by 12%. Overall, this study highlights the importance of dltB deletion and signal peptide optimization in enhancing extracellular protein production.
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Dan Xue, Zhuo Shang, Ethan A. Older, Zheng Zhong, Conor Pulliam, Kyle Peter, Mitzi Nagarkatti, Prakash Nagarkatti, Yong-Xin Li, Jie Li
Summary: Class III lanthipeptides are an underexplored subclass of lanthipeptides with potentially diverse chemical diversity and important biological activity. In this study, a biosynthetic pathway refactoring and heterologous overexpression strategy was designed and constructed to overcome the hurdles of silent BGCs and low-yielding pathways in the discovery of class III lanthipeptides from Firmicutes. This strategy enabled the discovery of new class III lanthipeptides bearing rare N,N-dimethylations and increased antibacterial activity.
ACS CHEMICAL BIOLOGY
(2023)
Article
Biochemical Research Methods
Stefano Grasso, Valentina Dabene, Margriet M. W. B. Hendriks, Priscilla Zwartjens, Rene Pellaux, Martin Held, Sven Panke, Jan Maarten van Dijl, Andreas Meyer, Tjeerd van Rij
Summary: The passage of proteins across biological membranes via the Sec pathway is a universally conserved process with critical functions. Estimating the impact of physicochemical signal peptide features on protein secretion levels was achieved through a high-throughput assay and machine learning model. The results allow for quantifying feature importance and predicting protein secretion levels, providing a versatile tool for signal peptide design and evaluation.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Genyu Wang, Zhijun Guo, Xueqian Zhang, Hao Wu, XiuMei Bai, Hailiang Zhang, Richa Hu, Shaoliang Han, Yuanxiang Pang, Zi'ang Gao, Lili Yan, Cuiying Huang, Le Zhang, Chunli Pan, Xuelian Liu
Summary: Listeria monocytogenes is a food-borne pathogen. Pediocin, a bacteriocin naturally produced by Pediococcus acidilacticus and Lactobacillus plantarum, has anti-listeria activity. In this study, researchers were able to express active pediocin/PapA in heterologous hosts without the assistance of accessory proteins. They also successfully purified the PA1 protein.
MICROBIAL CELL FACTORIES
(2022)
Article
Biotechnology & Applied Microbiology
Yuanyuan Xia, Lukasz Peplowski, Zhongyi Cheng, Tianyi Wang, Zhongmei Liu, Wenjing Cui, Michihiko Kobayashi, Zhemin Zhou
BIOTECHNOLOGY AND BIOENGINEERING
(2019)
Article
Agriculture, Multidisciplinary
Bo Pang, Li Zhou, Wenjing Cui, Zhongmei Liu, Shengmin Zhou, Jun Xu, Zhemin Zhou
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2019)
Article
Biochemistry & Molecular Biology
Li Zhou, Yuxi Wang, Hui Liu, Laichuang Han, Weimiao Zhang, Wenjing Cui, Zhongmei Liu, Zhemin Zhou
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
(2019)
Article
Biotechnology & Applied Microbiology
Laichuang Han, Qiaoqing Chen, Qiao Lin, Jintao Cheng, Li Zhou, Zhongmei Liu, Junling Guo, Linpei Zhang, Wenjing Cui, Zhemin Zhou
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2020)
Article
Biotechnology & Applied Microbiology
Laichuang Han, Wenjing Cui, Qiao Lin, Qiaoqing Chen, Feiya Suo, Ke Ma, Yang Wang, Wenliang Hao, Zhongyi Cheng, Zhemin Zhou
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2020)
Article
Biochemistry & Molecular Biology
Qiaoqing Chen, Wenjing Cui, Zhemin Zhou, Laichuang Han
Summary: In this study, the molecular dynamics of ligand and RNA binding by the anti-terminator protein GlpP were investigated through molecular docking, MD simulation, and alanine scanning mutagenesis. Results identified a ligand binding pocket and observed conformational changes in the RNA binding region, crucially involving residue interactions. Additionally, mutants were generated to confirm these findings, deepening understanding of GlpP as a potential regulator for gene expression in metabolic engineering and synthetic biology applications.
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
(2021)
Article
Chemistry, Physical
Yufeng Liu, Laichuang Han, Zhongyi Cheng, Zhongmei Liu, Zhemin Zhou
Article
Agriculture, Multidisciplinary
Li Zhou, Yuxi Wang, Laichuang Han, Qin Wang, Haili Liu, Ping Cheng, Ruoxuan Li, Xuecong Guo, Zhemin Zhou
Summary: This study successfully produced patchoulol in E. coli, achieving high production levels through various strategies, including engineering of patchoulol synthase and genetic manipulation of competitive pathways.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2021)
Article
Biochemical Research Methods
Wenjing Cui, Qiao Lin, Ruichun Hu, Laichuang Han, Zhongyi Cheng, Linpei Zhang, Zhemin Zhou
Summary: Efficient transcription termination using intrinsic terminators is crucial for cell fitness in bacteria. A data-driven and in silico-assisted (DISA) design framework was established to engineer minimal intrinsic terminators (MITs) using key features identified in natural intrinsic terminators. The MITs showed high termination efficiency in a wide range of bacteria compared to natural intrinsic terminators, and their efficiency could be flexibly tuned by modulating the length of the U-tract.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Agriculture, Multidisciplinary
Yuanyuan Xia, Wenwen Guo, Laichuang Han, Wei Shen, Xianzhong Chen, Haiquan Yang
Summary: This study obtained mutants with greatly improved activity and stability by site-directed mutagenesis of three potential residues of FruSG. The affinity and catalytic efficiency of mutant C43N were significantly enhanced, and its thermal and pH stability were also improved. The findings suggest that these mutations can enhance the performance of fructosyltransferase in industrial fructo-oligosaccharide production.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Biochemical Research Methods
Laichuang Han, Xinyue Liu, Zhongyi Cheng, Wenjing Cui, Junling Guo, Jian Yin, Zhemin Zhou
Summary: In this study, a desensitized niacin biosensor was constructed and used to develop a high-throughput screening platform for nitrile metabolism-related enzymes. Mutants with improved activity were obtained and showed broad applicability.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Li Zhou, Ying Zhu, Zhongzhe Yuan, Guangqing Liu, Zijin Sun, Shiyu Du, He Liu, Yating Li, Haili Liu, Zhemin Zhou
Summary: Efficient microbial strain was developed for 2-ketoisovalerate synthesis, achieving high titer, productivity, and yield through metabolic engineering strategies including overexpression of key enzymes, tuning NADPH supply, recycling NADH, and degrading pyruvate dehydrogenase. Modification of acetolactate synthase activity and expression was done to reduce by-products. These strategies proved effective in addressing redox imbalance and improving 2-ketoisovalerate production.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2022)