The influence of medium composition on the microbial secretory production of hydroxyalkanoate oligomers.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General and Applied Microbiology Pub Date : 2021-10-22 Epub Date: 2021-05-01 DOI:10.2323/jgam.2020.09.002
Shoji Mizuno, Tetsuo Sakurai, Mikito Nabasama, Kyouhei Kawakami, Ayaka Hiroe, Seiichi Taguchi, Takeharu Tsuge
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引用次数: 1

Abstract

With the aid of a chain transfer (CT) reaction, hydroxyalkanoate (HA) oligomers can be secreted by recombinant Escherichia coli carrying the gene encoding a lactate-polymerizing enzyme (PhaC1PsSTQK) in Luria-Bertani (LB) medium supplemented with a carbon source and CT agent. In this study, HA oligomers were produced through microbial secretion using a mineral-based medium instead of LB medium, and the impact of medium composition on HA oligomer secretion was investigated. The focused targets were medium composition and NaCl concentration related to osmotic conditions. It was observed that 4.21 g/L HA oligomer was secreted by recombinant E. coli in LB medium, but the amount secreted in the mineral-based modified R (MR) medium was negligible. However, when the MR medium was supplemented with 5 g/L yeast extract, 3.75 g/L HA oligomer was secreted. This can be accounted for by the enhanced expression and activity of PhaC1PsSTQK upon supplementation with growth-activated nutrients as supplementation with yeast extract also promoted cell growth and intracellular growth-associated polymer accumulation. Furthermore, upon adding 10 g/L NaCl to the yeast extract-supplemented MR medium, HA oligomer secretion increased to 6.86 g/L, implying that NaCl-induced osmotic pressure promotes HA oligomer secretion. These findings may facilitate the secretory production of HA oligomers using an inexpensive medium.

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培养基组成对微生物分泌羟基烷酸酯低聚物的影响。
通过链转移(CT)反应,携带编码乳酸聚合酶(PhaC1PsSTQK)基因的重组大肠杆菌可以在添加碳源和CT剂的Luria-Bertani (LB)培养基中分泌羟基烷烃酸(HA)低聚物。本研究采用矿物基培养基代替LB培养基,通过微生物分泌产生HA低聚物,并研究了培养基组成对HA低聚物分泌的影响。重点研究了与渗透条件有关的介质组成和NaCl浓度。结果发现,重组大肠杆菌在LB培养基中可分泌4.21 g/L HA低聚物,而在矿物基改性R (MR)培养基中可忽略不计。而在MR培养基中添加5 g/L酵母提取物时,HA低聚物的分泌量为3.75 g/L。这可以通过补充生长激活营养素增强PhaC1PsSTQK的表达和活性来解释,因为补充酵母提取物也促进细胞生长和细胞内生长相关的聚合物积累。在酵母提取物添加10 g/L NaCl的MR培养基中,HA寡聚物的分泌量增加到6.86 g/L,说明NaCl诱导的渗透压促进了HA寡聚物的分泌。这些发现可能有助于使用廉价培养基分泌HA低聚物的生产。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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