Arai Sayuri, Hayashihara Kayoko, Kanamoto Yuki, Shimizu Kazunori, Hirokawa Yasutaka, Hanai Taizo, Murakami Akio, Honda Hiroyuki
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8603, Japan.
Kobe University Research Center for Inland Seas, Awaji, Hyogo, Japan.
Biotechnol Bioeng. 2017 Aug;114(8):1771-1778. doi: 10.1002/bit.26307. Epub 2017 May 18.
Enhancement of alcohol tolerance in microorganisms is an important strategy for improving bioalcohol productivity. Although cyanobacteria can be used as a promising biocatalyst to produce various alcohols directly from CO , low productivity, and low tolerance against alcohols are the main issues to be resolved. Nevertheless, to date, a mutant with increasing alcohol tolerance has rarely been reported. In this study, we attempted to select isopropanol (IPA)-tolerant mutants of Synechococcus elongatus PCC 7942 using UV-C-induced random mutagenesis, followed by enrichment of the tolerant candidates in medium containing 10 g/L IPA and screening of the cells with a high growth rate in the single cell culture system in liquid medium containing 10 g/L IPA. We successfully acquired the most tolerant strain, SY1043, which maintains the ability to grow in medium containing 30 g/L IPA. The photosynthetic oxygen-evolving activities of SY1043 were almost same in cells after 72 h incubation under light with or without 10 g/L IPA, while the activity of the wild-type was remarkably decreased after the incubation with IPA. SY1043 also showed higher tolerance to ethanol, 1-butanol, isobutanol, and 1-pentanol than the wild type. These results suggest that SY1043 would be a promising candidate to improve alcohol production using cyanobacteria. Biotechnol. Bioeng. 2017;114: 1771-1778. © 2017 Wiley Periodicals, Inc.
提高微生物对酒精的耐受性是提高生物酒精产量的重要策略。尽管蓝藻可作为一种有前景的生物催化剂直接从二氧化碳生产各种酒精,但低产量和对酒精的低耐受性是需要解决的主要问题。然而,迄今为止,很少有关于酒精耐受性增强的突变体的报道。在本研究中,我们尝试使用紫外线C诱导的随机诱变来筛选耐异丙醇(IPA)的聚球藻PCC 7942突变体,随后在含有10 g/L IPA的培养基中富集耐受性候选菌株,并在含有10 g/L IPA的液体培养基的单细胞培养系统中筛选具有高生长速率的细胞。我们成功获得了耐受性最强的菌株SY1043,它在含有30 g/L IPA的培养基中仍保持生长能力。在有或没有10 g/L IPA的光照下培养72小时后,SY1043细胞的光合放氧活性几乎相同,而野生型细胞在与IPA孵育后活性显著下降。SY1043对乙醇、1-丁醇、异丁醇和1-戊醇的耐受性也高于野生型。这些结果表明,SY1043有望成为利用蓝藻提高酒精产量的候选菌株。《生物技术与生物工程》2017年;114: 1771 - 1778。© 2017威利期刊公司