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Transcriptional and enzymatic profiling of Pleurotus ostreatus laccase genes in submerged- and solid-state fermentation cultures

机译:深层和固态发酵培养物中平菇漆酶基因的转录和酶促分析

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摘要

The genome of the white rot basidiomycete Pleurotus ostreatus includes 12 phenol oxidase (laccase) genes. In this study, we examined their expression profiles in different fungal strains under different culture conditions (submerged and solid cultures) and in the presence of a wheat straw extract, which was used as an inducer of the laccase gene family. We used a reverse transcription-quantitative PCR (RT-qPCR)-based approach and focused on determining the reaction parameters (in particular, the reference gene set for the normalization and reaction efficiency determinations) used to achieve an accurate estimation of the relative gene expression values. The results suggested that (i) laccase gene transcription is upregulated in the induced submerged fermentation (iSmF) cultures but downregulated in the solid fermentation (SSF) cultures, (ii) the Lacc2 and Lacc10 genes are the main sources of laccase activity in the iSmF cultures upon induction with water-soluble wheat straw extracts, and (iii) an additional, as-yet-uncharacterized activity (Unk1) is specifically induced in SSF cultures that complements the activity of Lacc2 and Lacc10. Moreover, both the enzymatic laccase activities and the Lacc gene family transcription profiles greatly differ between closely related strains. These differences can be targeted for biotechnological breeding programs for enzyme production in submerged fermentation reactors.
机译:白色腐烂担子菌侧耳侧耳的基因组包括12个酚氧化酶(漆酶)基因。在这项研究中,我们检查了它们在不同培养条件(淹没和固体培养)下和存在麦草提取物(用作漆酶基因家族的诱导剂)的情况下在不同真菌菌株中的表达谱。我们使用了基于逆转录定量PCR(RT-qPCR)的方法,并着重于确定用于准确估计相对基因表达的反应参数(尤其是用于标准化和反应效率确定的参考基因集)价值观。结果表明(i)漆酶基因转录在诱导的深层发酵(iSmF)培养中被上调,而在固体发酵(SSF)培养中被下调;(ii)Lacc2和Lacc10基因是iSmF中漆酶活性的主要来源水溶性小麦秸秆提取物诱导后培养,(iii)在SSF培养物中特异诱导了另外一种尚未表征的活性(Unk1),可补充Lacc2和Lacc10的活性。此外,在密切相关的菌株之间,酶的漆酶活性和Lacc基因家族的转录谱都大大不同。这些差异可以作为在水下发酵反应器中生产酶的生物技术育种计划的目标。

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