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首页> 外文期刊>Microbial Cell Factories >Engineering marine fungi for conversion of d -galacturonic acid to mucic acid
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Engineering marine fungi for conversion of d -galacturonic acid to mucic acid

机译：用于将D-GalactuRonic酸转化为粘性酸的工程海洋真菌

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Two marine fungi, a Trichoderma sp. and a Coniochaeta sp., which can grow on d-galacturonic acid and pectin, were selected as hosts to engineer for mucic acid production, assessing the suitability of marine fungi for production of platform chemicals. The pathway for biotechnologcial production of mucic (galactaric) acid from d-galacturonic acid is simple and requires minimal modification of the genome, optimally one deletion and one insertion. d-Galacturonic acid, the main component of pectin, is a potential substrate for bioconversion, since pectin-rich waste is abundant. Trichoderma sp. LF328 and Coniochaeta sp. MF729 were engineered using CRISPR-Cas9 to oxidize d-galacturonic acid to mucic acid, disrupting the endogenous pathway for d-galacturonic acid catabolism when inserting a gene encoding bacterial uronate dehydrogenase. The uronate dehydrogenase was expressed under control of a synthetic expression system, which fucntioned in both marine strains. The marine Trichoderma transformants produced 25?g L?1 mucic acid from d-galacturonic acid in equimolar amounts: the yield was 1.0 to 1.1?g mucic acid [g d-galacturonic acid utilized]?1. d-Xylose and lactose were the preferred co-substrates. The engineered marine Trichoderma sp. was more productive than the best Trichoderma reesei strain (D-161646) described in the literature to date, that had been engineered to produce mucic acid. With marine Coniochaeta transformants, d-glucose was the preferred co-substrate, but the highest yield was 0.82?g?g?1: a portion of d-galacturonic acid was still metabolized. Coniochaeta sp. transformants produced adequate pectinases to produce mucic acid from pectin, but Trichoderma sp. transformants did not. Both marine species were successfully engineered using CRISPR-Cas9 and the synthetic expression system was functional in both species. Although Coniochaeta sp. transformants produced mucic acid directly from pectin, the metabolism of d-galacturonic acid was not completely disrupted and mucic acid amounts were low. The d-galacturonic pathway was completely disrupted in the transformants of the marine Trichoderma sp., which produced more mucic acid than a previously constructed T. reesei mucic acid producing strain, when grown under similar conditions. This demonstrated that marine fungi may be useful as production organisms, not only for native enzymes or bioactive compounds, but also for other compounds.
机译：两个海洋真菌，一个trichoderma sp。和一个Coniochaeta sp。，它可以在D-半乳糖酸和果胶上生长，选择为粘性酸生产的宿主，评估海洋真菌用于生产平台化学品的适用性。来自D-半乳糖醛酸的生物技术生产的生物技术生产的途径简单，需要最佳地改性基因组，最佳地进行一次缺失和一种插入。果胶的主要成分D-半乳糖醛酸是生物转化的潜在基质，因为富含果胶的废物是丰富的。 Trichoderma sp。 lf328和coniochaeta sp。使用CRISPR-CAS9设计MF729以将D-半乳糖醛酸氧化成粘性酸，当插入编码细菌尿酸脱氢酶的基因时，破坏D-半乳糖醛酸分解代谢的内源途径。在合成表达系统的控制下表达尿酸脱氢酶，其在两个海洋菌株中粘合。海洋richoderma转化体在等摩尔量中产生25μm-g 1粘蛋白，来自D-半溶栓的量：产率为1.0至1.1·g粘酸[使用]α1。 D-木糖和乳糖是优选的共衬底。工程型海洋Trichoderma sp。比文献中描述的最佳Trichoderma Reesei菌株（D-161646）迄今为止的最佳Trichoderma Reesei菌株（D-161646）更高效，这已经被设计成生产粘性酸。通过海洋Coniochaeta转化体，D-葡萄糖是优选的副​​衬底，但最高收率为0.82Ω·克··Δ1：仍代谢D-半乳糖醛酸的一部分。 Coniochaeta sp。转化体产生了足够的果胶酶，以产生来自果胶的阳性酸，但是Trichoderma Sp。转化体没有。使用CRISPR-CAS9成功设计了两个海洋物种，并且合成表达系统在这两个物种中都是功能的。虽然coniochaeta sp。转化体直接产生粘蛋白，D-半乳糖醛酸的代谢不完全破坏，粘酸量低。 D-半乳糖型途径在海洋richoderma Sp的转化体中完全破坏，其在相似条件下生长时产生比先前构造的T.reesei粘酸产生菌株更多的粘性酸。这证明海洋真菌可能是有用的生产生物，而不仅适用于原生酶或生物活性化合物，而且还可用于其他化合物。

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