Systems Microbiology and Biomanufacturing
·
2025
·
IF 2.9
Li Zihan,Zhao Guihong,Zhang Dezhi,Tang Yaqun,Liu Geer,Wang Xiaoyuan
2′-Fucosyllactose is the most abundant human milk oligosaccharides and one of the three essential nutrients for infant growth. Corynebacterium glutamicum is one of the most common industrial fermentation bacteria but cannot synthesize 2′-fucosyllactose. In this study, C. glutamicum ATCC13032 was engineered for 2′-fucosyllactose production from fucose and lactose. The gene futC from Helicobacter pylori encoding α-1,2-fucosyltransferase was codon optimized and mutated at four amino acids (F40S/Q150H/C151R/Q239S). The modified gene futC and the gene fkp from Bacteroides thetaiotaomicron encoding fucokinase/GDP-fucose pyrophosphorylase were overexpressed in plasmid pEC and transformed into C. glutamicum , resulting in CW002. CW002 did not synthesize 2′-fucosyllactose possibly because the substrates fucose and/or lactose did not pass through the cell membrane. Therefore, the gene lacY encoding lactose permease and the gene fucP encoding fucose permease from Escherichia coli were overexpressed in plasmid pXTuf and transformed into CW002, resulting in CW006. CW006 did synthesize 2′-fucosyllactose as expected. It is interesting that the production of 2′-fucosyllactose was decreased or stopped when the expression combination of these four genes was changed, suggesting that the expression levels of the four genes in CW006 might have to well balanced. C. glutamicum CW006 produced 2.07 g/L 2′-fucosyllactose in a 2.4 L bioreactor.
研究领域:
微生物学
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生物工程
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分子生物学
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食品科学
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DOI
高影响因子
Foods
·
2024
·Vol. 13
·期 9
·1386
·
IF 4.7
Li-Hao Wang,Wen-Hui Qu,Ya-Nan Xu,Song-Gang Xia,Qian-Qian Xue,Xiao-Ming Jiang,Hong-Ying Liu,Chang-Hu Xue,Yun-Qi Wen
The traditional fermentation process of soy sauce employs a hyperhaline model and has a long fermentation period. A hyperhaline model can improve fermentation speed, but easily leads to the contamination of miscellaneous bacteria and fermentation failure. In this study, after the conventional koji and moromi fermentation, the fermentation broth was pasteurized and diluted, and then inoculated with three selected microorganisms includingCorynebacterium glutamicum,Corynebacterium ammoniagenes, andLactiplantibacillus plantarumfor secondary fermentation. During this ten-day fermentation, the pH, free amino acids, organic acids, nucleotide acids, fatty acids, and volatile compounds were analyzed. The fermentation group inoculated withC. glutamicumaccumulated the high content of amino acid nitrogen of 0.92 g/100 mL and glutamic acid of 509.4 mg/100 mL. TheC. ammoniagenesgroup andL. plantarumgroup were rich in nucleotide and organic acid, respectively. The fermentation group inoculated with three microorganisms exhibited the best sensory attributes, showing the potential to develop a suitable fermentation method. The brewing speed of the proposed process in this study was faster than that of the traditional method, and the umami substances could be significantly accumulated in this low-salt fermented model (7%w/vNaCl). This study provides a reference for the low-salt and rapid fermentation of seasoning.
研究领域:
食品科学
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微生物学
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DOI
PubMed
高影响因子
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
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2025
·
IF 1.4
Chanyu Wu,Xiaohan Wang,Xincan Li,Hexian Li,Qiuyue Peng,Xiaoxin Niu,Yutong Wu,Zhiying Wang,Zuoyong Zhou
Corynebacterium pseudotuberculosis , a zoonotic intracellular bacteria, is responsible for abscesses and pyogranuloma formation of the infected host, which is essentially a chronic inflammatory response. Tripartite motif-containing protein 21 (TRIM21) negatively regulates pro-inflammatory cytokines production during C . pseudotuberculosis infection, the mechanism of which remains unclear. This study found that C . pseudotuberculosis infection in macrophages induced phosphorylation of IκB and p65. TRIM21 interacted with IκBα by PRY/SPRY domain, stabilizes IκBα and negatively regulates IκBα phosphorylation in macrophages during C. pseudotuberculosis infection. In addition, TRIM21 positively regulates the ubiquitination of IκBα via K48 linkage rather than K63 linkage in C. pseudotuberculosis -infected macrophages. In brief, our research confirmed that TRIM21 negatively regulates canonical NF-κB activation by interacting with IκBα and decreasing IκBα phosphorylation in macrophages during C. pseudotuberculosis infection. Preventing inflammation induced by C. pseudotuberculosis infection through regulation of the NF-κB pathway is a potential way to control this pathogen.
研究领域:
免疫学
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微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2018
·Vol. 102
·期 14
·6039-6046
·
IF 3.34
Tigu Fitsum,Zhang Junli,Liu Guoxia,Cai Zhen,Li Yin
d -Pantothenic acid (vitamin B 5 ) has wide applications in the feed, food, chemical, and pharmaceutical industries. Its biological production routes which employ pantothenate synthetase (PS) as the key enzyme are attractive since they avoid the tedious and time-consuming optical resolution process. However, little data is available on the activity and kinetics of this enzyme, hampering the rational selection of an efficient enzyme for the biological production of d -pantothenic acid. In this study, six phylogenetically distant PS-encoding genes, from Escherichia coli , Corynebacterium glutamicum , Bacillus subtilis , Bacillus thuringiensis , Bacillus cereus , and Enterobacter cloacae , were expressed in E. coli . The PS from C. glutamicum exhibited a specific activity of 205.1 U/mg and a turnover number of 127.6 s -1 , which to our best knowledge are the highest values ever reported. The addition of substrates ( d -pantoic acid and β-alanine) to the E. coli strain harboring this enzyme during the early log phase of fermentation resulted in the production of 97.1 g/L of d -pantothenic acid within 32 h, corresponding to a conversion yield of 99.1% and a productivity of 3.0 g/L/h. To the best of our knowledge, this is the highest productivity reported to date.
研究领域:
生物化学
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微生物学
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工业生物技术
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2025
·
IF 3.8
Ying-Ying Xu,Bin Zhang,Jie Bao
This study investigated Corynebacterium glutamicum for GABA production using corncob residue, a cheap carbon source, as carbohydrate feedstock instead of grains. Current fermentative production of γ-aminobutyric acid (GABA) mainly relies on starch and sugar feedstocks. Therefore, the use of cheap carbon sources can effectively reduce the production costs of GABA. This study validated the feasibility of an available non-food lignocellulose feedstock, corncob residue, for GABA fermentation. An engineered C. glutamicum was constructed by secretory expression of glutamate decarboxylase gadBmut and knock-out the GABA biodegradation gene. The fermentation parameters in shake flasks and bioreactors were optimized including the sugar concentration, nitrogen levels, cofactor levels and aeration rate, and fermentation mode. The fed-batch fermentation of the concentrated corncob residue hydrolysate achieved a record high GABA production of 93.15 g/L from lignocellulosic feedstock with a yield of 0.43 g/g glucose. The results provided technical support for the industrial production of GABA from low-cost lignocellulosic carbohydrates.
研究领域:
生物技术
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微生物学
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产品关联
DOI
高影响因子
JOURNAL OF BIOTECHNOLOGY
·
2021
·
IF 3.307
Jingbai Wen,Jie Bao
Fermentative production of γ-aminobutyric acid by the glutamate overproducing Corynebacterium glutamicum from cheap sugar feedstock is generally regarded as one of the most promising methods to reduce the production cost. However, the intracellularly expressed glutamate decarboxylase in C. glutamicum often showed feeble catalysis activity to convert glutamate into γ-aminobutyric acid. Here we tried to secretory express glutamate decarboxylase to achieve efficient extracellular decarboxylation of glutamate, thus improving the γ-aminobutyric acid production by C. glutamicum . We first tested glutamate decarboxylases from different sources, and the mutated glutamate decarboxylase GadBmut from E. coli with better catalytic performance was selected. Then, a signal peptide of the SEC translocation pathway directed the successful secretion of glutamate decarboxylase in C. glutamicum . The extracellular catalysis by secreted glutamate decarboxylase increased the γ-aminobutyric acid generation by three-fold, compared with intracellular catalysis. Enhancing glutamate decarboxylase expression and decreasing γ-aminobutyric acid degradation further increased γ-aminobutyric acid production by 39 %. The fed-batch fermentation of the engineered C. glutamicum strain reached the record high titer (77.6 ± 0.0 g /L), overall yield (0.44 ± 0.00 g/g glucose), and productivity (1.21 ± 0.00 g/L/h). This study demonstrated a unique design of extracellular catalysis for efficient γ-aminobutyric acid production by C. glutamicum .
研究领域:
微生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2025
·
IF 3.8
Ying-Ying Xu,Bin Zhang,Jie Bao
Industrial production of cadaverine primarily relies on whole-cell catalysis of lysine using engineered Escherichia coli as host cells. The growing interest in cadaverine as a monomer of biobased polyamides requires inexpensive, non-food carbohydrate feedstock for its bioproduction, rather than using food-derived sugars as feedstock. This study used corncob residue as the starting carbohydrate feedstock followed by enzymatic hydrolysis to obtain the sugars. Since E. coli was not tolerant to even minor inhibitor residues in corncob residue hydrolysate, the robust Corynebacterium glutamicum was used as the host bacterium followed by the metabolic modifications of secretory expression of lysine decarboxylase via Ncgl1289 and cgR_0949 , and cadaverine degradation pathway knockout. The final whole-cell catalysis of C. glutamicum recombinant using corncob hydrolysate as carbohydrate feedstock achieved a record-high 78.19 g/L of cadaverine with a conversion yield of 91%.
研究领域:
工业微生物学
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生物工程
·
代谢工程
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产品关联
DOI
高影响因子
ELECTROPHORESIS
·
2019
·Vol. 40
·期 20
·2665-2671
·
IF 2.754
Wei Wang,Ruiguang Bai,Huimin Zhang,Xiaoyu Cai
Corynebacterium glutamicum ( C. glutamicum ) is a well-known workhorse for the industrial production of amino acids. Different carbon, nitrogen, and sulfur source may force the bacterium to produce specific metabolites. In this work, a method of high-speed MEKC with LIF detection was developed to rapidly analyze the amino acid metabolites released by C. glutamicum , which is fed with different culture mediums. Corynebacterium glutamicum was cultured in microbial fuel cells to monitor its metabolism process and collect its metabolites. In the CE system, a microliter-scale sample reservoir was designed and applied to perform tiny volume sample injection. With the assistance of microwave, the derivatization time for amino acids with FITC was greatly shortened to 6 min. Under the optimized condition, the eight candidate amino acids of metabolites could be rapidly separated within 2 min. The whole analysis process for real samples, from sampling to determination, could be shortened to less than 10 min. The results showed that C. glutamicum could produce additional l -lysine and l -valine as the metabolites when fed with glucose and l -methionine, respectively. The method proved that culture mediums used to feed C. glutamicum had great effect on the bacterium's metabolites.
研究领域:
微生物学
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生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
·
2024
·
IF 4.4
Hui-Zhong Sun,Si-Yu Wei,Qiu-Man Xu,Wei Shang,Qing Li,Jing-Sheng Cheng,Ying-Jin Yuan
Polymyxin B, produced by Paenibacillus polymyxa, is used as the last line of defense clinically. In this study, exogenous mixture of precursor amino acids increased the level and proportion of polymyxin B1 in the total of polymyxin B analogs of P. polymyxa CJX518-AC (PPAC) from 0.15 g/L and 61.8 % to 0.33 g/L and 79.9 %, respectively. The co-culture of strain PPAC and recombinant Corynebacterium glutamicum -leu01, which produces high levels of threonine, leucine, and isoleucine, increased polymyxin B1 production to 0.64 g/L. When strains PPAC and C. glu -leu01 simultaneously inoculated into an optimized medium with 20 g/L peptone, polymyxin B1 production was increased to 0.97 g/L. Furthermore, the polymyxin B1 production in the co-culture of strains PPAC and C. glu -leu01 increased to 2.21 g/L after optimized inoculation ratios and fermentation medium with 60 g/L peptone. This study provides a new strategy to improve polymyxin B1 production.
研究领域:
微生物学
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生物工程
·
药学
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产品关联
DOI
PubMed
高影响因子
Systems Microbiology and Biomanufacturing
·
2024
·
IF 0
Huang Yu,Li Hedan,Zhao Guihong,Hu Xiaoqing,Wang Xiaoyuan
The production of L-alanine was enhanced in Corynebacterium glutamicum ATCC13869 through metabolic engineering of the biosynthesis pathways of L-alanine and fatty acids. Strains Δ fasB , Δ fasBR , Δ fasB Δ pks13 and Δ fasBR Δ pks13 were constructed and exhibited increased L-alanine yields up to 17.29 g/L. Different from Δ fasB mutant constructed from C. glutamicum ATCC13032 in which L-glutamate production accumulated, the muatnt Δ fasB constructed from C. glutamicum ATCC13869 in this study significantly produce L-alanine without L-glutamate accumulation. Transcriptional level analysis revealed that the knockout of fasB upregulated the expression levels of the genes related to L-alanine synthesis but downregulated those associated with fatty acid synthesis, confirming the redirection of metabolic flux from fatty acid synthesis to L-alanine synthesis in these strains. L-alanine productions were further enhanced in strains Δ fasB and Δ fasBR through the combinatorial expression of heterologous genes Bacillus subtilis alaD encoding alanine dehydrogenase and Escherichia coli alaE encoding alanine export protein, and the yields reached 55.21 g/L and 54.95 g/L, respectively. Finally, 69.9 g/L L-alanine was obtained in Δ fasB /pJYW-5- alaDE after 60 h of fermentation by supplementing glucose. Our data indicate that disrupting the fatty acid biosynthesis could redirect metabolic flux towards L-alanine biosynthesis. These results provide a new strategy for increasing the production of L-alanine in C. glutamicum . Graphical Abstract
研究领域:
微生物学
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生物工程
·
分子生物学
✓ 已验证
DOI
ACS Catalysis
·
2025
·
IF 13.1
Mengkai Hu,Teng Bao,Zhen Qin,Qiang Wang,Hengwei Zhang,Yujue Wang,Jiajia You,Zhenglian Xue,Rongzhen Zhang,Shang-Tian Yang,Xian Zhang,Zhiming Rao
Surface-displayed whole-cell biocatalysis offers significant advantages over traditional intracellular enzyme-based methods but faces challenges in complex multienzyme processes and low efficiency. We present Scaffold-Mediated Assembly Regulation Tool (SMART), a programmable platform for dynamic control of multienzyme biocatalysts on Corynebacterium glutamicum surfaces. SMART integrates orthogonal cross-linked scaffolds with the twin Catcher/Tag pairs (SpyCatcher/SpyTag and SnoopCatcher/SnoopTag), controllable molecular ratios, and high loading capacity, offering user-defined functionality for enzyme assembly. We optimized SMART through computational modeling and experimental validation and created bifunctional scaffolds and long tandem repeats for optimal enzyme loading. The versatility of SMART was demonstrated by codisplaying in a two enzyme sequential cascade with tunable stoichiometry, significantly improving the conversion of maltodextrin to trehalose. Furthermore, we integrated a Bxb1 recombinase-based state machine (RSM) genetic circuit, allowing temporal regulation of enzyme and scaffold expression within a single cell. Finally, the system’s applicability was showcased in isomaltulose production from low-cost sucrose. SMART design represents a significant advancement in surface-displayed biocatalysis, offering a programmable platform for complex multienzyme reactions with potential applications in various industrially relevant biocatalysts.
研究领域:
合成生物学
·
生物化学
·
代谢工程
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Shijin Wu,Xiaojing Ma,Hongde Yan
The asymmetric reduction of α , β -unsaturated compounds conjugated with electron-withdrawing group by ene-reductases (ERs) is a valuable method for the synthesis of enantiopure chiral compounds. This study introduced an ER from Corynebacterium casei ( Cc ER) which was heterologously expressed in Escherichia coli BL21(DE3), and the purified recombinant Cc ER was characterized for its biocatalytic properties. Cc ER exhibited the highest specific activity at 40 °C and pH 6.5, and showcased appreciable stability below 40 °C over a pH range of 6.0–7.0. The enzyme displayed high resistance to methanol. Cc ER accepted NADH or NADPH as a cofactor and exhibited a broad substrate spectrum towards α , β -unsaturated compounds. It achieved complete conversion of 2-cyclohexen-1-one and good performance for stereoselective reduction of ( R )-carvone (conversion 98 %, diastereoselectivity 96 %). This study highlights the robustness and potential of Cc ER.
研究领域:
生物化学
·
生物技术
·
酶学
·
有机化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）