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从苦荞麦种子中分离筛选产大量芦丁降解酶的菌株。

Isolation and screening of strains producing high amounts of rutin degrading enzymes from Fagopyrum tataricum seeds.

机构信息

College of Life and Basic Science, Sichuan Agricultural University, Yaan, Sichuan, People's Republic of China.

出版信息

J Basic Microbiol. 2013 Feb;53(2):181-7. doi: 10.1002/jobm.201100337. Epub 2012 May 14.

Abstract

The rutin degrading enzyme (RDE) was isolated and purified from tartary buckwheat seeds. The RDE was purified about 11.34-fold and its final yield was 3.5%, which was very low, due to our purification strategy of giving priority to purity over yield. The RDE molecular weight was estimated to be about 60 kDa. When rutin was used as substrate, an optimal enzyme activity was seen at around pH 5.0 and 40 °C. Strains isolation strategy characterized by the use of rutin as sole carbon source in enrichment cultures was used to isolate RDE-producing strains. Then the active strains were identified by morphology characterization and 18s rDNA-ITS (Internal Transcribed Spacer) gene sequencing. Three isolates coded as B3, W2, Y2 were successfully isolated from fusty Fagopyrum tataricum flour cultures. Strain B3 possessed the highest unit activity among these three strains, and its total activity reached up to 171.0 Unit. The active isolate (B3) could be assigned to Penicillium farinosum. When the Penicillium farinosum strains were added to tartary buckwheat flour cultures at pH 5.0, 30 °C after 5 days fermentation, the quercetin production raised up to 1.78 mg/l, almost 5.1 times higher than the fermentation without the above active strains. Hence, a new approach was available to utilize microorganism-aided fermentation for effective quercetin extraction from Fagopyrum tataricum seeds.

摘要

芦丁降解酶(RDE)从苦荞种子中分离和纯化。由于我们的纯化策略优先考虑纯度而不是产量,因此 RDE 被纯化了约 11.34 倍,最终得率为 3.5%,非常低。RDE 的分子量估计约为 60 kDa。当芦丁作为底物时,在 pH 5.0 和 40°C 左右观察到最佳酶活性。采用以芦丁为唯一碳源的富集培养物来分离 RDE 产生菌的菌株分离策略。然后通过形态特征和 18s rDNA-ITS(内部转录间隔区)基因测序来鉴定活性菌株。从发霉的苦荞面粉培养物中成功分离出编码为 B3、W2、Y2 的 3 个分离株。在这 3 个分离株中,B3 菌株的单位活性最高,总活性达到 171.0 单位。活性分离株(B3)可归为青霉属。当青霉属菌株在 pH 5.0、30°C 下添加到苦荞面粉培养物中,发酵 5 天后,槲皮素的产量增加到 1.78mg/L,几乎比没有上述活性菌株的发酵高出 5.1 倍。因此,利用微生物辅助发酵从苦荞种子中有效提取槲皮素的新方法成为可能。

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