Development of efficient 5-ketogluconate production system by Gluconobacter japonicus
Development of efficient 5-ketogluconate production system by Gluconobacter japonicus
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日本葡萄糖酸杆菌高效生产5-酮葡萄糖酸系统的开发
DOI：
10.1007/s00253-022-12242-0
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Yakushi T.
中科院分区：
文献类型：
--
作者：
Kataoka N;Naoki K;Ano Y;Matsushita K;Yakushi T.
5-酮葡糖酸盐（5 KGA）是合成酒石酸盐的前体，酒石酸盐是一种在几个行业中使用的有价值的化合物。在先前的研究中，缺乏两种膜葡糖酸2-糖苷酶的日本葡糖酸杆菌NBRC 3271突变株D2显示从葡萄糖和葡糖酸的混合物产生5 KGA而不是2-酮葡糖酸。在这项研究中，我们的目的是开发一个有效的5 KGA生产系统使用G。japonicusD 2为亲本。D2在罐式发酵罐培养中从葡萄糖产生5 KGA;然而，在培养的后期阶段，5 KGA水平降低。为了增加D2产生5 KGA的潜力，修饰了与5 KGA和葡萄糖酸盐利用相关的细胞质代谢;从D2中分别删除编码5 KGA还原酶和葡糖激酶的gno和gntK基因，产生D4。与D2相比，在D4中观察到5 KGA产量的提高，但在培养结束时仍有大量的葡萄糖酸盐，导致0.83 mol（mol葡萄糖）-1的产率不满意。葡萄糖酸盐转化为5 KGA是由吡咯喹啉醌（PQQ）依赖性甘油脱氢酶（GLDH）催化的，GLDH很容易通过释放PQQ和钙离子形成脱辅基酶。因此，研究了培养基中添加CaCl 2对D4产生5 KGA的影响。我们证明了1 mM CaCl 2的添加对PQQ-GLDH对葡萄糖酸盐的活性的维持产生了积极影响，从而提高了5 KGA的产量，产量达到0.97 mol（mol葡萄糖）− 1。关键点·用葡萄糖酸盐开发了一种有效的5 KGA生产系统。·删除gno和gntK基因阻断了5 KGA和葡萄糖酸盐的催化作用。添加1 mM CaCl 2有效地提高了葡萄糖向5 KGA的转化率。
5-Ketogluconate (5KGA) is a precursor for synthesizing tartrate, a valuable compound used in several industries. In a previous study,Gluconobacter japonicusNBRC 3271 mutant strain D2, which lacks two membranous gluconate 2-dehydrogenases, was shown to produce 5KGA but not 2-ketogluconate from a mixture of glucose and gluconate. In this study, we aimed to develop an efficient 5KGA production system usingG. japonicusD2 as the parental strain. D2 produced 5KGA from glucose in a jar fermentor culture; however, 5KGA levels were reduced during the late phase of cultivation. To increase the potential of D2 for 5KGA production, the cytoplasmic metabolism related to the utilization of 5KGA and gluconate was modified; thegnoandgntKgenes encoding 5KGA reductase and gluconokinase, respectively, were deleted from D2, generating D4. Improved 5KGA production was observed in D4 compared to that in D2, but a significant amount of gluconate remained at the end of cultivation, leading to an unsatisfied yield of 0.83 mol (mol glucose)−1. The conversion of gluconate to 5KGA is catalyzed by pyrroloquinoline quinone (PQQ)-dependent glycerol dehydrogenase (GLDH), which easily forms an apoenzyme by releasing PQQ and calcium ions. Thus, the effects of CaCl2addition to the culture medium on 5KGA production by D4 were investigated. We demonstrated that 1 mM CaCl2addition positively affected the maintenance of the PQQ-GLDH activity toward gluconate and consequently enhanced 5KGA production, and the yield reached 0.97 mol (mol glucose)−1.Key points•An efficient 5KGA production system was developed with Gluconobacter japonicus.•Deleting the gno and gntK genes blocked the catabolism of 5KGA and gluconate.•The addition of 1 mM CaCl2efficiently improved the conversion of glucose to 5KGA.
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影响因子：
4.4
作者：
Ittipon Saichana;D. Moonmangmee;O. Adachi;K. Matsushita;H. Toyama
通讯作者：
Ittipon Saichana;D. Moonmangmee;O. Adachi;K. Matsushita;H. Toyama
影响因子：
5
作者：
Elfari, M;Ha, SW;Görisch, H
通讯作者：
Görisch, H
影响因子：
3.5
作者：
INOUE, H;NOJIMA, H;OKAYAMA, H
通讯作者：
OKAYAMA, H
工业葡糖杆菌膜结合甘油脱氢酶的溶解、纯化和性质
DOI：
10.1080/00021369.1985.10866858
发表时间：
1985
期刊：
Agricultural and biological chemistry
影响因子：
--
作者：
M. Ameyama;E. Shinagawa;K. Matsushita;O. Adachi
通讯作者：
O. Adachi
日本葡萄糖杆菌NBRC 3271膜结合葡萄糖酸脱氢酶的研究
DOI：
--
发表时间：
2019
期刊：
影响因子：
--
作者：
片岡 尚也;大田 諒子;Natsaran Saichana;松谷 峰之介;外山 博英;藥師 寿治;松下 一信
通讯作者：
松下 一信