ENZYME AND MICROBIAL TECHNOLOGY
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2023
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IF 3.4
Shuangshuang Duan,Yonghua Chen,Guodong Wang,Zebin Li,Shitong Dong,Yingshuai Wu,Yuanwei Wang,Chunling Ma,Ruiming Wang
D -Allose is a rare cis-caprose with a wide range of physiological functions, which has a wide range of applications in medicine, food, and other industries. L -Rhamnose isomerase ( L -Rhi) is the earliest enzyme found to catalyze the production of D -allose from D -psicose. This catalyst has a high conversion rate, but its specificity for substrates is limited; thus, it cannot fulfill the requirements of industrial production of D -allose. In this study, L -Rhi derived from Bacillus subtilis was employed as the research subject, and D -psicose as the conversion substrate. Two mutant libraries were constructed through alanine scanning, saturation mutation, and rational design based on the analysis of the secondary structure , tertiary structure , and interactions with ligands of the enzyme. The yield of D -allose produced by these mutants was assessed; it was found that the conversion rate of mutant D325M to D -allose was increased by 55.73 %, and the D325S improved by 15.34 %, while mutant W184H increased by 10.37 % at 55 °C, respectively. According to modeling analysis, manganese (Mn 2+ ) had no significant effect on the production of D -psicose from D -psicose by L -Rhi. The results of molecular dynamics simulation demonstrated that the mutants W184H, D325M, and D325S had more stable protein structures while binding with the substrate D -psicose, as evidenced by its root mean square deviation (RMSD), root mean square fluctuation (RMSF), and binding free energy values. It was more conducive to binding D -psicose and facilitating its conversion to D -allose, providing the basis for the production of D -allose.
研究领域:
生物化学
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酶学
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生物技术
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结构生物学
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产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
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2022
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IF 3.705
Lei Wang,Kecai Chen,Peng Zheng,Xiang Huo,Fei Liao,Liping Zhu,Meirong Hu,Yong Tao
D -Psicose, a new-generation sugar substitute, has been enzymatically synthesized through D -fructose isomerization. However, isomerization often causes low yields due to unfavorable thermodynamic equilibria, which limited its further industrial application. In this study, we present a redox-driven multi-enzyme cascade, two-step biotransformation system to produce D -psicose from D -fructose. Compared to D -fructose isomerization, this method has a maximized theoretical conversion rate of 100%. D -Psicose-3-epimerase from Clostridiales (CBDPE), ribitol 2-dehydrogenase from Providencia alcalifaciens (PRDH), and formate dehydrogenase from Starkeya (SFDH) were co-expressed in Escherichia coli in the first step to produce D -allitol from D -fructose. Afterward, NADH oxidase from Streptococcus pyogenes (SPNOX), and ribitol 2-dehydrogenase from Rubrivivax sp. (RSRDH) were co-expressed in E. coli to oxidize D -allitol into D -psicose in the second step. The two-step biotransformation system was optimized to maximize the D -fructose-to- D -psicose conversion rate (up to 90%), corresponding to a concentration of 450 mM. This study suggests that this redox-driven multi-enzyme cascade strategy through a sugar-to-alcohol-to-sugar pathway has the advantage of great application for enhanced production of D -psicose and other rare sugars.
研究领域:
生物化学
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生物技术
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酶学
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代谢工程
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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2023
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IF 3.1
Yibo Guo,Ke Shen,Xinshuai Zhang,Hua Huang
l -threonate is the metabolite of vitamin C, while d -erythronate is the metabolite of N -acetyl- d -glucosamine, the nutritional supplement for joint health. They are widely distributed in the environment and human biofluids. Nevertheless, the catabolisms of l -threonate and d -erythronate are sparsely reported. Here we explored the functional diversity of an acid sugar kinase family (Pfam families PF07005-PF17042), and discovered a novel 2-oxo-tetronate kinase. The conserved genome neighborhood of the 2-oxo-tetronate kinase encodes members of class-II fructose-bisphosphate aldolase family (F_bP_aldolase, PF01116) and a dehydrogenase family (PF03446-PF14833). Instructed by this analysis, we experimentally verified that these enzymes are capable of degrading l -threonate into dihydroxyacetone phosphate (DHAP) in Arthrobacter sp. ZBG10, Clostridium scindens ATCC 35704, and Pseudonocardia dioxanivorans ATCC 55486. Meanwhile, a convergent catabolic pathway for d -erythronate was characterized in P . dioxanivorans ATCC 55486. Moreover, the phylogenetic distribution analysis indicates that the biological range of the identified l -threonate and d -erythronate catabolic pathways appears to extend mostly to members of the Actinomycetota , Cyanobacteriota , Bacillota , Pseudomonadota , and Bacteroidota phyla.
研究领域:
微生物学
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生物化学
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分子生物学
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基因组学
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代谢
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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）