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与运动发酵单胞菌的葡萄糖酸内酯酶相关的工业鲁棒性。

Industrial robustness linked to the gluconolactonase from Zymomonas mobilis.

机构信息

School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Kensington, 2052, Australia.

Department of Microbiology, Pukyong National University, Busan, 608-737, Republic of Korea.

出版信息

Appl Microbiol Biotechnol. 2017 Jun;101(12):5089-5099. doi: 10.1007/s00253-017-8248-y. Epub 2017 Mar 24.

Abstract

The physiological characteristics and the potential gluconolactone production of the gluconolactonase-deficient strain, Zymomonas mobilis ZM4 gnlΔ, were investigated via growth inhibitory assay and biotransformation of glucose and fructose into gluconolactone and sorbitol, respectively. The results of ethanol fermentation studies performed in the presence of high concentration of glucose (>200 g l) under fermentative or aerobic conditions indicated that a significant reduction of volumetric ethanol productivity from the strain of ZM4 gnlΔ was noticeable due to the reduced rates of specific growth, sugar uptake, and biomass yield as compared with those of the parental strain ZM4. The biotransformation prepared at pH 6.0 using the permeabilized cell indicated that gluconic acid from ZM4 gnlΔ was still produced as a major product (67 g l) together with sorbitol (65 g l) rather than gluconolactone after 24 h. Only small amount of gluconolactone was transiently overproduced up to 9 g l, but at the end of biotransformation, all gluconolactone were oxidized into gluconic acid. This indicated that autolysis of gluconolactone at the pH led to such results despite under gluconolactonase inactivation conditions. The physiological characteristics of ZM4 gnlΔ was further investigated under various stress conditions, including suboptimal pH (3.56.0), temperature (2540 °C), and presence of growth inhibitory molecules including hydrogen peroxide, ethanol, acetic acid, furfural, and so forth. The results indicated that ZM4 gnlΔ was more susceptible at high glucose concentration, low pH of 3.5, and high temperature of 40 °C and in the presence of 4 mM HO comparing with ZM4. Therefore, the results were evident that gluconolactonase in Z. mobilis contributed to industrial robustness and anti-stress regulation.

摘要

通过生长抑制试验以及将葡萄糖和果糖分别转化为葡萄糖酸内酯和山梨醇来研究缺乏葡萄糖酸内酯酶的菌株,运动发酵单胞菌 ZM4 gnlΔ 的生理特性和潜在的葡萄糖酸内酯生产能力。在发酵或需氧条件下,当存在高浓度葡萄糖(>200 g l)时进行的乙醇发酵研究结果表明,与亲本菌株 ZM4 相比,由于比生长速率、糖摄取率和生物量产率降低,ZN4 gnlΔ 菌株的体积乙醇生产率显著降低。在 pH 6.0 下使用透性细胞进行的生物转化表明,与葡萄糖酸内酯相比,ZM4 gnlΔ 仍产生主要产物葡萄糖酸(67 g l)和山梨醇(65 g l)。只有少量的葡萄糖酸内酯在 24 小时内暂时过量生产,达到 9 g l,但在生物转化结束时,所有的葡萄糖酸内酯都被氧化成葡萄糖酸。这表明尽管在缺乏葡萄糖酸内酯酶的条件下,pH 下的自溶作用导致了这种结果。在各种胁迫条件下,包括非最佳 pH(3.56.0)、温度(2540°C)以及存在生长抑制分子,包括过氧化氢、乙醇、乙酸、糠醛等,进一步研究了 ZM4 gnlΔ 的生理特性。结果表明,与 ZM4 相比,ZN4 gnlΔ 在高葡萄糖浓度、低 pH 值 3.5 和高温 40°C 以及存在 4 mM HO 时更为敏感。因此,结果表明运动发酵单胞菌中的葡萄糖酸内酯酶有助于工业鲁棒性和抗应激调节。

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