识别 Pseudogluconobacter saccharoketogenes D-葡萄糖酸生产的途径
Tetsuya Ito1, Hisaharu Masaki1, Koki Fujita1
1ENSUIKO Sugar Refining Co., Ltd., Tokyo, 103-0012, Japan.
Applied biochemistry and biotechnology
|July 13, 2023
概括
伪葡萄糖菌 (Pseudogluconobacter saccharoketogenes) 通过两个途径产生葡萄糖酸. 这项研究确定了关键中间体D-glucaraldehyde和L-guluronic acid,澄清了葡萄糖酸生物合成中的酶作用.
科学领域:
- 微生物学 微生物学
- 生物化学 生物化学
- 代谢工程是代谢工程.
背景情况:
- 众所周知,Pseudogluconobacter saccharoketogenes可以从D-葡萄糖中产生葡萄糖酸.
- 存在两个主要的生物合成途径,通过D-葡萄酸或D-葡萄酸进行.
- 酒精脱酶,化脱酶和葡萄糖酸脱酶的特定作用,以及D-葡萄糖和L-氨酸中间体的参与,尚未完全阐明.
研究的目的:
- 在葡萄糖酸生产中识别和描述拟议的中间体D-葡萄糖化物和L-氨酸.
- 阐明酒精脱酶,化脱酶和葡萄糖酸脱酶在这些途径中的作用.
- 为了确定参与葡萄糖酸生物合成的关键酶的基质特异性.
主要方法:
- 净化和识别D-葡萄糖化物和L-氨酸.
- 酶测试以确定酒精脱酶,化脱酶和葡萄糖酸脱酶在中间体和氧化产品上的基质特异性.
- 使用P. saccharoketogenes Rh47-3.3的休息细胞进行大规模发酵 (30-L).
主要成果:
- D-glucaraldehyde和L-guluronic acid被成功净化,并被确定为中间体.
- 酒精脱酶表现出广泛的基质特异性,而化脱酶则具有有限的基质氧化.
- 大规模生产从D-葡萄糖中获得60.3mol% (7.0g/L) 的D-葡萄糖酸,从D-葡萄糖酸获得78.6mol% (22.5g/L) 的D-葡萄糖酸.
结论:
- 该研究证实了D-glucaraldehyde和L-guluronic acid作为P. saccharoketogenes在葡萄糖酸生产中的中间体的作用.
- 酶表征提供了关于酒精,化物和葡萄糖酸盐脱酶在生物合成途径中的特定功能的见解.
- 优化条件显示了高效的D-葡萄糖酸生产,突出了工业应用的潜力.
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