中国农业科学 ›› 2016, Vol. 49 ›› Issue (3): 573-580.doi: 10.3864/j.issn.0578-1752.2016.03.015

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

豆粕与发酵豆粕中主要抗营养因子调查分析

杨玉娟1,2,姚怡莎1,秦玉昌2,邱静3,李军国1,李俊1,谷旭1   

  1. 1中国农业科学院饲料研究所,北京100081
    2农业部食物与营养发展研究所,北京100081
    3中国农业科学院农业质量标准与检测技术研究所,北京100081
  • 收稿日期:2015-01-07 出版日期:2016-02-01 发布日期:2016-02-01
  • 通讯作者: 谷旭,Tel:010-82106069;E-mail:guxu@caas.cn
  • 作者简介:杨玉娟,E-mail:yyj464636994@163.com。姚怡莎:E-mail:yaoyisha521@163.com。杨玉娟和姚怡莎为同等贡献作者。
  • 基金资助:
    国家自然科学基金(21407176)、国家公益性行业(农业)科研专项(201203015)、现代农业产业技术体系北京市家禽创新团队专项资金项目(CZ1108)

Investigation and Analysis of Main AFN in Soybean Meal and Fermented Soybean Meal

YANG Yu-juan1,2, YAO Yi-sha1, QIN Yu-chang2, QIU Jing3, LI Jun-guo1, LI Jun1, GU Xu1   

  1. 1Feed Research Institute, Chinese Academy of Agricultural Science, Beijing 100081
    2Institute of Food and Nutrition Development, Ministry of Agriculture, Beijing 100081
    3Institute of Quality Standard and Testing Technology for Agro-products, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2015-01-07 Online:2016-02-01 Published:2016-02-01

摘要: 【目的】豆粕是动物饲料的主要原料,但其含多种抗营养因子(anti-nutritional factors, ANF),阻碍营养成分的消化、吸收和利用,从而影响动物的生长发育和健康。研究表明豆粕经微生物发酵可有效地降低抗营养因子含量。但由于发酵工艺、发酵菌种、豆粕本身的因素,不同生产厂家的豆粕及发酵豆粕中各抗营养因子含量差别较大,现有研究中也少有关于二者中抗营养因子水平的研究报道。为此,抽取了市售的65批次豆粕和54批次发酵豆粕,对6抗营养因子:大豆球蛋白、β-伴大豆球蛋白、胰蛋白酶抑制因子、棉籽糖、水苏糖、脲酶进行分析测定,以了解饲料行业使用的豆粕及发酵豆粕中的抗营养因子含量。【方法】用ELISA法(enzyme-linked immuno sorbent assay)对样品中的大豆球蛋白、β-伴大豆球蛋白、胰蛋白酶抑制因子含量进行测定,其分析方法和操作要求均与所购ELISA试剂盒的说明相一致,主要过程为:样品前处理、加样、洗板、加酶标试剂、显色、终止。棉籽糖和水苏糖的检测采用高效液相色谱(high performance liquid chromatography, HPLC)检测微波提取的棉籽糖和水苏糖。脲酶分析参照国标方法:加入尿素缓冲液后恒温水浴,一定时间后加入盐酸溶液停止反应后冷却,清洗试管内容物,以氢氧化钠标准溶液滴定至pH4.7后根据体积计算得出脲酶活性。【结果】调查分析后发现:豆粕和发酵豆粕中的大豆球蛋白平均含量分别为129.3、54.7 mg·g-1,发酵后大豆球蛋白平均含量降低了57.7%,根据百分位数法对数据进行统计分析,得出豆粕和发酵豆粕中的大豆球蛋白正常值范围分别为58.9—P90(177.3 mg·g-1)、ND—P90(109.4 mg·g-1)。豆粕中的β-伴大豆球蛋白平均含量为102.2 mg·g-1,而发酵豆粕中的β-伴大豆球蛋白为37.6 mg·g-1,相比豆粕降低了63.2%,使用相同的数据统计方法判定二者中β-伴大豆球蛋白含量正常值范围分别为42.8—P85(147.2 mg·g-1)和ND—P85(61.8 mg·g-1)。胰蛋白酶抑制因子在豆粕和发酵豆粕中平均含量分别为18.4 mg·g-17.5 mg·g-1,发酵处理使其含量下降了59.1%,同时得出豆粕及发酵豆粕胰蛋白抑制因子含量正常值范围分别在ND—P80(28.6 mg·g-1)、ND—P80(9.9 mg·g-1)之间。豆粕和发酵豆粕中的棉籽糖平均含量分别为11.02、1.93 mg·g-1,发酵豆粕比豆粕减少了82.5%,豆粕和发酵豆粕中棉籽糖的正常值范围分别在ND—P90 (13.79 mg·g-1)、ND—P90(4.65 mg·g-1)之间。豆粕中水苏糖的平均含量为29.70 mg·g-1,而发酵豆粕中水苏糖的平均含量为5.19 mg·g-1,发酵后水苏糖含量降低了82.5%,同时水苏糖的正常值范围分别在ND—P85 (33.29 mg·g-1)、ND—P85(11.58 mg·g-1)之间;豆粕中脲酶含量正常值范围为ND—P97(0.40 U·g-1),发酵豆粕脲酶未检出。综上得出,发酵豆粕的抗营养因子含量与豆粕相比有不同程度的减少。【结论】在分析调查的基础上得出了现行市售豆粕及发酵豆粕主要抗营养因子的含量范围。本调查分析为饲料加工工艺的进一步优化提供数据支撑,同时能够对养殖企业选择豆粕及发酵豆粕作为饲料原材料起到一定的理论指导作用。

关键词: 豆粕, 发酵豆粕, 大豆抗营养因子

Abstract: 【Objective】Soybean meal is the main raw material of feed. However, various anti-nutritional factors (ANF) in soybean meal hinder the digestion, absorption and utilization for nutrients, which would have negative effects on animal growth and health. Studies have shown that soybean meal fermented by microorganism could decrease the undesirable effect of ANF. While, fermentation strains, soybean species, and manufacturers may have an effect in fermentation process. Moreover, seldom reports referred to the levels of ANF in soybean meal and fermented soybean meal. Based on this, 65 batches of available soybean meal and 54 batches of fermented soybean meal were collected in this study, followed by analysis of 6 kinds of ANF, including glycinin, β-conglycinin, trypsin inhibitors, raffinose, stachyose and urease, in order to investigate the actual levels of ANF in soybean meal and fermented soybean meal.【Method】In this study, glycinin, β-conglycinin and trypsin inhibitors were analyzed by Enzyme-link Immunosorbent Assay (ELISA) method. Experimental procedures were conducted according to instructions which were sample pretreatment, plus, washing, adding enzymatic reagent, color reaction, and termination. The analysis method of raffinose and stachyose was HPLC after extracted by microwave. Analysis of urease refers to GB/T 8622-2006, and titration was utilized for analysis of urease activity.【Result】The average concentration of glycinin in soybean meal (129.3 mg·g-1) is 57.7% higher than fermented soybean meal (54.7 mg·g-1). The percentile method was used for analysis. The normal values in soybean meal and fermented soybean were 58.9-177.3 mg·g-1 and ND (Not Detected)-109.4 mg·g-1, respectively. Moreover, the fermented soybean meal (37.6 mg·g-1) is 63.2% lower than the concentration of β-conglycinin in soybean meal (102.2 mg·g-1). The normal values in soybean meal and fermented soybean were 42.8-147.2 mg·g-1 and ND-61.8 mg·g-1, respectively. The average concentration of trypsin inhibitors in soybean meal (18.4 mg·g-1) is 59.1% higher than fermented soybean meal (7.5 mg·g-1). In addition, the normal values of trypsin inhibitor in soybean meal and fermented soybean meal were ND-28.9 mg·g-1 and ND-9.9 mg·g-1. The average concentration of raffinose in soybean meal and fermented soybean meal were 11.02 mg·g-1 and 1.93 mg·g-1, with the former is 80% higher. The normal values of raffinose in soybean meal and fermented soybean meal were ND-13.79 mg·g-1 and ND-4.65 mg·g-1. The average value of stachyose average in soybean meal (29.70 mg·g-1) is higher than in fermented soybean meal (5.19 mg·g-1). The normal values of stachyose are ND-33.29 mg·g-1, ND-11.58 mg·g-1, respectively. The normal values of urease in soybean meal was in the range of ND to 0.40 U·g-1, while the activity value of urease of fermented soybean meal was under the detection limit.  It was concluded that the concentrations of ANF in fermented soybean meal have decreased differently compared with soybean meal.【Conclusion】This research provided data support for further optimization of feed processing, and could act as a guide for breeding companies to choose higher quality of soybean meal and fermented soybean meal as feed raw materials.

Key words: soybean meal, fermented soybean meal, AFN