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Construction of Recombinant Producers of Enzyme Preparations for Feed Production with an Expression System Based on Penicillium verruculosum Fungus
Applied Biochemistry and Microbiology ( IF 1.0 ) Pub Date : 2020-12-01 , DOI: 10.1134/s0003683820080104
A. P. Sinitsyn , O. G. Korotkova , E. A. Rubtsova , O. A. Sinitsyna , E. G. Kondrat’eva , A. S. Sereda , I. N. Zorov , A. M. Rozhkova

Abstract

Producer strains have been created with an expression system based on the recipient strain Penicillium verruculosum 537 (ΔniaD) and the cellobiohydrolase-1 gene promoter. These strains made it possible to obtain new enzyme preparations (EPs) for feed production that were characterized by a high content of endodepolymerases (endo-β-1,4-glucanases and endo-β-1,4-xylanase), which degrade nonstarch polysaccharides (NSPs) of grain cereals used as a feed component. The proportion of endodepolymerases in the new EPs was 44–68% of the total protein content (as compared to 15% in the recipient-based preparation), whereas the percentage of exodepolymerases (cellobiohydrolases) fell to 8–26% as compared to 60% in the recipient strain-based product. The new EPs have a high specific activity of endodepolymerases, which was 1.8–4.6 times greater than that in the EP obtained from the recipient strain, and the specific activity of exodepolymerases decreased by 2.0–3.7 times. The new EPs demonstrated endoglucanase and xylanase activities in a wide range of pH and temperature, including the physiological values of these parameters (pH 3.0 and 7.0, and t 37–38°C). They were characterized by high endodepolymerase stability under the action of digestive proteases (pepsin and trypsin). The EPs also retained their endoglucanase and xylanase activities during feed granulation (80°C). The xylanase contained in the new EPs was not affected by proteinaceous inhibitors from cereals. Experimental batches of EPs were made by the Agroferment enzyme-producing plant and were tested in broiler and piglet feeding. The addition of the new commercial EPs to the animals’ diets increased their live weight, reduced feed and protein intake, and decreased the metabolic energy per unit of body weight, improving digestibility and assimilability of feed nutrients, especially raw fiber and NSP.



中文翻译:

基于疣状青霉菌表达系统的饲料生产用酶制剂重组生产者的构建

摘要

已经使用基于受体菌株Vericulium verruculosum 537(ΔniaD)和纤维二糖水解酶-1基因启动子。这些菌株使得获得用于饲料生产的新酶制剂成为可能,这些酶制剂的特点是内切聚合酶(内切-β-1,4-葡聚糖酶和内切-β-1,4-木聚糖酶)含量高,这些酶会降解非淀粉。谷物谷物的多糖(NSP)用作饲料成分。新EP中内切聚合酶的比例为总蛋白质含量的44–68%(相比,以受体为基础的制剂中为15%),而外切聚合酶(纤维二糖水解酶)的比例为60–60%,降至8–26% %的基于受体菌株的产品。新的EP具有高的内切聚合酶比活性,比从受体菌株获得的EP高1.8-4.6倍,而外聚合酶的比活性降低了2.0-3.7倍。t 37–38°C)。它们的特征是在消化蛋白酶(胃蛋白酶和胰蛋白酶)的作用下具有高内切聚合酶稳定性。EP在饲料颗粒化过程中(80°C)还保留了其内切葡聚糖酶和木聚糖酶活性。新EP中包含的木聚糖酶不受谷物蛋白质抑制剂的影响。由农用酶生产工厂生产实验批次的EP,并在肉鸡和仔猪饲喂中进行测试。在动物的饮食中添加新的商业EP会增加它们的活体重,减少饲料和蛋白质的摄入,并降低每单位体重的代谢能,从而提高饲料营养物质(特别是粗纤维和NSP)的消化率和消化率。

更新日期:2020-12-01
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