Hao Wei, Tian Pengjiao, Zheng Mingli, Wang Huili, Xu Chuncheng
Department of Agricultural Engineering, College of Engineering, China Agricultural University, Beijing 100083, China.
State Key Laboratory of Feed Microbial Engineering, Beijing Da Bei Nong Science and Technology Group Co., Ltd., Beijing, China.
Asian-Australas J Anim Sci. 2020 Jan 1;33(1):100-110. doi: 10.5713/ajas.18.0933. Epub 2019 Apr 15.
The objective of this study was to isolate proteolytic microorganisms and evaluate their effects on proteolysis in total mixed ration (TMR) silages of soybean curd residue.
TMRs were formulated with soybean curd residue, alfalfa or Leymus chinensis hay, corn meal, soybean meal, a vitamin-mineral supplement, and salt in a ratio of 25.0:40.0:30.0:4.0:0.5:0.5, respectively, on a basis of dry matter (DM). The microbial proteinases during ensiling were characterized, the dominate strains associated with proteolysis were identified, and their enzymatic characterization were evaluated in alfalfa (A-TMR) and Leymus chinensis (L-TMR) TMR silages containing soybean curd residue.
Both A-TMR and L-TMR silages were well preserved, with low pH and high lactic acid concentrations. The aerobic bacteria and yeast counts in both TMR silages decreased to about 105 cfu g-1 FM (Fresh matter) and below the detection limit, respectively. The lactic acid bacteria count increased to 109 cfu g-1 FM. The total microbial proteinases activities reached their maximums during the early ensiling stage and then reduced in both TMR silages with fermentation prolonged. Metalloproteinase was the main proteinase when the total proteinases activities reached their maximums, and when ensiling terminated, metallo and serine proteinases played equally important parts in proteolysis in both TMR silages. Strains in the genera Curtobacterium and Paenibacillus were identified as the most dominant proteolytic bacteria in A-TMR and L-TMR, respectively, and both their proteinases were mainly with metalloproteinase characteristics. In the latter ensiling phase, Enterococcus faecium strains became the major sources of proteolytic enzymes in both TMR silages. Their proteinases were mainly of metallo and serine proteinases classes in this experiment.
Proteolytic aerobic bacteria were substituted by proteolytic lactic acid bacteria during ensiling, and the microbial serine and metallo proteinases in these strains played leading roles in proteolysis in TMR silages.
本研究旨在分离蛋白水解微生物,并评估其对豆腐渣全混合日粮(TMR)青贮饲料中蛋白水解的影响。
以干物质(DM)为基础,将豆腐渣、苜蓿或羊草干草、玉米粉、豆粕、维生素 - 矿物质预混料和盐按25.0:40.0:30.0:4.0:0.5:0.5的比例配制TMR。对青贮过程中的微生物蛋白酶进行了表征,鉴定了与蛋白水解相关的优势菌株,并在含豆腐渣的苜蓿(A - TMR)和羊草(L - TMR)TMR青贮饲料中评估了它们的酶学特性。
A - TMR和L - TMR青贮饲料均保存良好,pH值低,乳酸浓度高。两种TMR青贮饲料中的需氧菌和酵母菌数量分别降至约105 cfu g-1鲜物质(FM)和低于检测限。乳酸菌数量增加到109 cfu g-1 FM。随着发酵时间延长,两种TMR青贮饲料中的总微生物蛋白酶活性在青贮初期达到最大值,然后降低。当总蛋白酶活性达到最大值时,金属蛋白酶是主要的蛋白酶,青贮结束时,金属蛋白酶和丝氨酸蛋白酶在两种TMR青贮饲料的蛋白水解中发挥同等重要的作用。分别鉴定出短小杆菌属和芽孢杆菌属的菌株为A - TMR和L - TMR中最主要的蛋白水解细菌,它们的蛋白酶主要具有金属蛋白酶特性。在青贮后期,粪肠球菌菌株成为两种TMR青贮饲料中蛋白水解酶的主要来源。在本实验中,它们的蛋白酶主要属于金属蛋白酶和丝氨酸蛋白酶类。
青贮过程中蛋白水解需氧菌被蛋白水解乳酸菌取代,这些菌株中的微生物丝氨酸和金属蛋白酶在TMR青贮饲料的蛋白水解中起主导作用。