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H2O2 as a candidate bottleneck for MnP activity during cultivation of Agaricus bisporus in compost

机译:H2O2作为堆肥双孢蘑菇培养过程中MnP活性的候选瓶颈

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摘要

Degradation of lignin by fungi enhances availability of cellulose and hemicellulose in plant waste and thereby increases the amount of carbon source available to these microorganisms. The button mushroom Agaricus bisporus degrades only about half of the lignin in compost and about 40% of the carbohydrates remain unutilized during mushroom cultivation. Here it was assessed whether over-expression of the manganese peroxidase gene mnp1 improves lignin degradation and, as a consequence, carbohydrate breakdown by A. bisporus. Transformants expressing mnp1 under the control of actin regulatory sequences produced MnP activity in malt extract medium, while the parental strain A15 did not. MnP activity was increased 0.3- and 3-fold at casing and after the 2nd flush of a semi-commercial cultivation, respectively, when compared to strain A15. Pyrolysis-GC–MS showed that overexpression of MnP decreased phenylmethane and phenylethane type lignin relative to the phenylpropane type after the 2nd flush. However, it neither affected the syringyl/guaiacyl derived residue ratio nor the ratio of oxidized to non-oxidized lignin residues. Moreover, the carbohydrate content and accessibility was not affected in compost. Notably, the capacity of compost extract to consume the MnP co-factor H2O2 was 4- to 8-fold higher than its production. This may well explain why over-expression of mnp1 did not improve carbohydrate degradation in compost. In fact, availability of H2O2 may limit lignin degradation by wild-type A. bisporus.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0424-z) contains supplementary material, which is available to authorized users.
机译:真菌对木质素的降解提高了植物废料中纤维素和半纤维素的利用率,从而增加了这些微生物可利用的碳源量。纽扣蘑菇蘑菇双孢蘑菇仅降解堆肥中约一半的木质素,而约40%的碳水化合物在蘑菇栽培期间仍未利用。在此评估了锰过氧化物酶基因mnp1的过表达是否改善了木质素降解,并因此改善了双孢曲霉对碳水化合物的降解作用。在肌动蛋白调节序列控制下表达mnp1的转化子在麦芽提取培养基中产生MnP活性,而亲本菌株A15则没有。与菌株A15相比,在半套管栽培和第2次冲洗后,MnP活性分别提高了0.3倍和3倍。热解-GC-MS表明,相对于第二次冲洗后的苯基丙烷类型,MnP的过表达降低了苯基甲烷和苯基乙烷型木质素。然而,它既不影响丁香基/愈创木酰基衍生的残基比率,也不影响氧化的与未氧化的木质素残基的比率。此外,堆肥中的碳水化合物含量和可及性不受影响。值得注意的是,堆肥提取物消耗MnP辅因子H2O2的能力比其生产高4至8倍。这可以很好地解释为什么mnp1的过表达不能改善堆肥中碳水化合物的降解。实际上,H2O2的可用性可能会限制野生型双孢曲霉降解木质素。电子补充材料本文的在线版本(doi:10.1186 / s13568-017-0424-z)包含补充材料,授权用户可以使用。

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