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Zhang, Liaoyuan,Singh, Raushan,D, Sivakumar,Guo, Zewang,Li, Jiahuan,Chen, Fanbing,He, Yuanzhi,Guan, Xiong,Kang, Yun Chan,Lee, Jung-Kul unknown 2018 Green Chemistry Vol. No.
<P>Upgrading ethanol to higher order alcohols is desired but difficult using current biotechnological methods. In this study, we designed a completely artificial reaction pathway for upgrading ethanol to acetoin, 2,3-butanediol, and 2-butanol in a cell-free bio-system composed of ethanol dehydrogenase, formolase, 2,3-butanediol dehydrogenase, diol dehydratase, and NADH oxidase. Under optimized conditions, acetoin, 2,3-butanediol, and 2-butanol were produced at 88.78%, 88.28%, and 27.25% of the theoretical yield from 100 mM ethanol, respectively. These results demonstrate that this artificial synthetic pathway is an environmentally-friendly novel approach for upgrading bio-ethanol to acetoin, 2,3-butanediol, and 2-butanol.</P>
( Liaoyuan Zhang ),( Zewang Guo ),( Huifang Gao ),( Xiaoqian Peng ),( Yongyu Li ),( Shujing Sun ),( Jung-kul Lee ),( Wenxiong Lin ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.12
Many plant-pathogenic bacteria are dependent on quorum sensing (QS) to evoke disease. In this study, the population of QS and quorum quenching (QQ) bacteria was analyzed in a consecutive monoculture system of Pseudostellaria heterophylla. The isolated QS strains were identified as Serratia marcescens with SwrIR-type QS system and exhibited a significant increase over the years of monoculture. Only one QQ strain was isolated from newly planted soil sample and was identified as Bacillus thuringiensis, which secreted lactonase to degrade QS signal molecules. Inoculation of S. marcescens to P. heterophylla root could rapidly cause wilt disease, which was alleviated by B. thuringiensis. Furthermore, the expression of lactonase encoded by the aiiA gene in S. marcescens resulted in reduction of its pathogenicity, implying that the toxic effect of S. marcescens on the seedlings was QS-regulated. Meanwhile, excess lactonase in S. marcescens led to reduction in antibacterial substances, exoenzymes, and swarming motility, which might contribute to pathogensis on the seedlings. Root exudates and root tuber extracts of P. heterophylla significantly promoted the growth of S. marcescens, whereas a slight increase of B. thuringiensis was observed in both samples. These results demonstrated that QS-regulated behaviors in S. marcescens mediated by root exudates played an important role in replanting diseases of P. heterophylla.
Jianan Sun,Liaoyuan Zhang,Ben Rao,Yunbin Han,Ju Chu,Jiawen Zhu,Yaling Shen,Dong-Zhi Wei 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.3
Enhanced acetoin production was carried out by Serratia marcescens H32. First, medium compositions were optimized statistically for shake flask fermentations to produce acetoin. Sucrose and corn steep liquor powder (CSLP) were identified as the most significant factors by Plackett–Burman design. The path of steepest ascent and response surface methodology were then employed to determine the optimal concentrations of the two factors. Acetoin yield was up to 41.5 g/L in flask fermentations using the optimized medium. Furthermore, the optimal medium was used to conduct fermentation experiments in a 3.7-L bioreactor. The influences of different agitation speeds on acetoin production were investigated. Based on a process analysis, a two-stage agitation speed control strategy was proposed, in which the agitation speed was controlled at 700 rpm during the first 8 h and then switched to 600 rpm. A relatively high acetoin concentration (44.9 g/L)and high acetoin productivity (1.73 g/L/h) were achieved by applying this strategy. Fed-batch fermentation based on the two-stage agitation speed control strategy was performed,and a maximum acetoin concentration of 60.5 g/L with productivity of 1.44 g/L/h was achieved. Enhanced acetoin production was carried out by Serratia marcescens H32. First, medium compositions were optimized statistically for shake flask fermentations to produce acetoin. Sucrose and corn steep liquor powder (CSLP) were identified as the most significant factors by Plackett–Burman design. The path of steepest ascent and response surface methodology were then employed to determine the optimal concentrations of the two factors. Acetoin yield was up to 41.5 g/L in flask fermentations using the optimized medium. Furthermore, the optimal medium was used to conduct fermentation experiments in a 3.7-L bioreactor. The influences of different agitation speeds on acetoin production were investigated. Based on a process analysis, a two-stage agitation speed control strategy was proposed, in which the agitation speed was controlled at 700 rpm during the first 8 h and then switched to 600 rpm. A relatively high acetoin concentration (44.9 g/L)and high acetoin productivity (1.73 g/L/h) were achieved by applying this strategy. Fed-batch fermentation based on the two-stage agitation speed control strategy was performed,and a maximum acetoin concentration of 60.5 g/L with productivity of 1.44 g/L/h was achieved.
( Tae Su Kim ),( Hyung Moo Jung ),( Sang Yong Kim ),( Liaoyuan Zhang ),( Jinglin Li ),( Sujan Sigdel ),( Ji Hyun Park ),( Jung Rim Haw ),( Jung Kul Lee ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.7
Acetate and lactate in growth media are detrimental to the production of Thermus maltogenicamylase (ThMA), a heterologous protein, as well as to the growth of recombinant Escherichia coli. Only 50 mM of acetate or 10 mM of lactate reduced 90% of specific ThMA activity. In this study, mutant E. coli strains blocked in the ackA-pta or ackA-pta and ldh pathways were created, characterized, and assessed for their culture performace in 300 L-scale fermentation. The ackApta and ldh double-mutant strain formed significantly less lactate and acetate, and produced a concomitant increase in the excretion of pyruvate (17.8 mM) under anaerobic conditions. The ackA-pta mutant strain accumulated significant acetate but had an approximately 2-fold increase in the formation of lactate. The ackA-pta and ldh double-mutant strain had superior overall performance in large-scale culture under suboptimal conditions, giving 67% higher cell density and 66% higher ThMA activity compared with those of the control strain. The doublemutant strain also achieved a 179% improvement in volumetric ThMA production.
( Zewang Guo ),( Xihua Zhao ),( Yuanzhi He ),( Tianxing Yang ),( Huifang Gao ),( Ganxin Li ),( Feixue Chen ),( Meijing Sun ),( Jung-kul Lee ),( Liaoyuan Zhang ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.1
Acetoin (AC) is a volatile platform compound with various potential industrial applications. AC contains two stereoisomeric forms: (3S)-AC and (3R)-AC. Optically pure AC is an important potential intermediate and widely used as a precursor to synthesize novel optically active materials. In this study, chiral (3R)-AC production from meso-2,3-butanediol (meso-2,3-BD) was obtained using recombinant Escherichia coli cells co-expressing meso-2,3-butanediol dehydrogenase (meso-2,3-BDH), NADH oxidase (NOX), and hemoglobin protein (VHB) from Serratia sp. T241, Lactobacillus brevis, and Vitreoscilla, respectively. The new biocatalyst of E. coli/pET-mbdh-nox-vgb was developed and the bioconversion conditions were optimized. Under the optimal conditions, 86.74 g/l of (3R)-AC with the productivity of 3.61 g/l/h and the stereoisomeric purity of 97.89% was achieved from 93.73 g/l meso-2,3-BD using the whole-cell biocatalyst. The yield and productivity were new records for (3R)-AC production. The results exhibit the industrial potential for (3R)-AC production via whole-cell biocatalysis.
( Wensong Jin ),( Hui Lin ),( Huifang Gao ),( Zewang Guo ),( Jiahuan Li ),( Quanming Xu ),( Shujing Sun ),( Kaihui Hu ),( Jung-kul Lee ),( Liaoyuan Zhang ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.4
N-acyl-homoserine lactone quorum sensing (AHL-QS) has been shown to regulate many physiological behaviors in Serratia marcescens MG1. In the current study, the effects of AHL-QS on the biosynthesis of acid and neutral products by S. marcescens MG1 and its isogenic ΔswrI with or without supplementing exogenous N-hexanoyl-L-homoserine lactone (C<sub>6</sub>-HSL) were systematically investigated. The results showed that swrI disruption resulted in rapid pH drops from 7.0 to 4.8, which could be restored to wild type by supplementing C<sub>6</sub>-HSL. Furthermore, fermentation product analysis indicated that ΔswrI could lead to obvious accumulation for acidogenesis products such as lactic acid and succinic acid, especially excess acetic acid (2.27 g/l) produced at the early stage of fermentation, whereas solventogenesis products by ΔswrI appeared to noticeably decrease by an approximate 30% for acetoin during 32-48 h and by an approximate 20% for 2,3-butanediol during 24-40 h, when compared to those by wild type. Interestingly, the excess acetic acid produced could be removed in an AHL-QS-independent manner. Subsequently, quantitative real-time PCR was used to determine the mRNA expression levels of genes responsible for acidogenesis and solventogenesis and showed consistent results with those of product synthesis. Finally, by close examination of promoter regions of the analyzed genes, four putative luxI box-like motifs were found upstream of genes encoding acetyl-CoA synthase, lactate dehydrogenase, α-acetolactate decarboxylase, and Lys-like regulator. The information from this study provides a novel insight into the roles played by AHL-QS in switching from acidogenesis to solventogenesis in S. marcescens MG1.
( S. V. Otari ),( S. H. Pawar ),( Sanjay K. S. Patel ),( Raushan K. Singh ),( Sang-yong Kim ),( Jai Hyo Lee ),( Liaoyuan Zhang ),( Jung-kul Lee ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.4
A novel approach to synthesize silver nanoparticles (AgNPs) using leaf extract of Canna edulis Ker-Gawl. (CELE) under ambient conditions is reported here. The as-prepared AgNPs were analyzed by UV-visible spectroscopy, transmission emission microscopy, X-ray diffraction, Fourier transform-infrared spectroscopy, energy-dispersive analysis of X-ray spectroscopy, zeta potential, and dynamic light scattering. The AgNPs showed excellent antimicrobial activity against various pathogens, including bacteria and various fungi. The biocompatibility of the AgNPs was analyzed in the L929 cell line using NRU and MTT assays. Acridine orange/ ethidium bromide staining was used to determine whether the AgNPs had necrotic or apoptotic effects on L929 cells. The concentration of AgNPs required for 50% inhibition of growth of mammalian cells is far more than that required for inhibition of pathogenic microorganisms. Thus, CELE is a candidate for the eco-friendly, clean, cost-effective, and nontoxic synthesis of AgNPs.
Production of Methanol from Methane by Encapsulated Methylosinus sporium
( Sanjay K. S. Patel ),( Jae-hoon Jeong ),( Sanjeet Mehariya ),( Sachin V. Otari ),( Bharat Madan ),( Jung Rim Haw ),( Jung-kul Lee ),( Liaoyuan Zhang ),( In-won Kim ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.12
Massive reserves of methane (CH<sub>4</sub>) remain unexplored as a feedstock for the production of liquid fuels and chemicals, mainly because of the lack of economically suitable and sustainable strategies for selective oxidation of CH4 to methanol. The present study demonstrates the bioconversion of CH<sub>4</sub> to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of a Type II methanotroph, Methylosinus sporium, was carried out using different encapsulation methods, employing sodium-alginate (Na-alginate) and silica gel. The encapsulated cells demonstrated higher stability for methanol production. The optimal pH, temperature, and agitation rate were determined to be pH 7.0, 30oC, and 175 rpm, respectively, using inoculum (1.5 mg of dry cell mass/ml) and 20% of CH<sub>4</sub> as a feed. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, the Na-alginate and silica gel encapsulated cells retained 61.8% and 51.6% of their initial efficiency for methanol production, respectively, in comparison with the efficiency of 11.5% observed in the case of free cells. These results suggest that encapsulation of methanotrophs is a promising approach to improve the stability of methanol production.