Properties of Two Novel Esterases Identified from Culture Supernatant of Penicillium purpurogenum Grown on Sugar Beet Pulp.

Insights in enzyme research Pub Date : 2016-01-01 Epub Date: 2016-12-12
Gabriela Oleas, Eduardo Callegari, Romina Sepulveda, Jaime Eyzaguirre
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Abstract

Background: The filamentous fungus Penicillium purpurogenum grows on a variety of natural carbon sources, such as sugar beet pulp, and secretes to the medium a large number of enzymes that degrade the carbohydrate components of lignocellulose. Sugar beet pulp is rich in pectin, and the purpose of this work is to identify novel esterases produced by the fungus, which may participate in pectin degradation.

Methods and findings: Partially purified culture supernatants of the fungus grown on sugar beet pulp were subjected to mass spectrometry analysis. Peptides thus identified, which may be part of potential esterases were probed against the proteins deduced from the fungal genome sequence. The cDNAs of two putative esterases identified were expressed in Pichia pastoris and their properties studied. One of these enzymes, named FAET, is a feruloyl esterase, while the other, PE, is classified as a pectin methyl esterase.

Conclusions: These findings add to our knowledge of the enzymology of pectin degradation by Penicillium purpurogenum, and define properties of two novel esterases acting on de-esterification of pectin. Their availability may be useful as tools for the study of pectin structure and degradation.

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甜菜果肉紫青霉培养上清中两种新型酯酶的性质研究。
背景:丝状真菌紫霉属(Penicillium purpurogenum)生长在多种天然碳源上,如甜菜浆,并向培养基分泌大量降解木质纤维素碳水化合物成分的酶。甜菜果肉中含有丰富的果胶,本研究的目的是鉴定由真菌产生的可能参与果胶降解的新型酯酶。方法和发现:用质谱法对甜菜果肉部分纯化的真菌培养上清液进行分析。由此鉴定出的肽可能是潜在酯酶的一部分,并与真菌基因组序列推断的蛋白质进行了探测。鉴定了两种可能的酯酶的cdna在毕赤酵母中表达,并研究了它们的性质。其中一种名为FAET的酶是阿魏酰酯酶,而另一种名为PE的酶则被归类为果胶甲基酯酶。结论:这些发现增加了我们对青霉菌降解果胶的酶学知识,并确定了两种作用于果胶去酯化的新型酯酶的性质。它们的可用性可以作为研究果胶结构和降解的有用工具。
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Properties of Two Novel Esterases Identified from Culture Supernatant of Penicillium purpurogenum Grown on Sugar Beet Pulp Properties of Two Novel Esterases Identified from Culture Supernatant of Penicillium purpurogenum Grown on Sugar Beet Pulp.
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