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31.
芽孢杆菌中性植酸酶基因的原核表达及酶学性质分析 总被引:1,自引:0,他引:1
植酸酶作为饲料添加剂能够有效提高动物对饲料中磷的利用率及减少粪便中磷排放对环境的污染,并降低植酸的抗营养作用。为了获得性能稳定的高活性植酸酶,采用PCR扩增芽孢杆菌(Bacillus sp.)中性植酸酶基因phyC(GenBank登录号:FJ986327)的成熟肽编码序列,将其克隆进原核表达载体pET-28a(+),并转化E.coli BL21(DE3)进行表达。在37℃条件下以0.5 mmol/L IPTG诱导4 h能够获得大量包涵体蛋白,在25℃条件下以0.5 mmol/L IPTG诱导6 h有利于可溶性蛋白的获得。利用Ni-NTA亲和层析柱纯化重组植酸酶产物,获得的中性植酸酶的部分酶学特性为:耐热性较好,最适反应温度55℃,在70℃处理10 min可保持20%以上的酶活性;耐酸、碱能力较强,最适pH 6.0~7.0,pH 5.5~9.0时能保持80%以上的酶活性,pH 5.0~10.0时处理60 min仍能保持70%以上的酶活性,在pH 2.0~4.0时能保持40%以上的酶活性。利用构建的切除芽孢杆菌中性植酸酶基因phyC信号肽编码序列的原核表达载体及优化的诱导表达条件,能够在大肠杆菌中高量表达性能稳定的芽孢杆菌中性植酸酶。 相似文献
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《The Journal of Applied Poultry Research》2014,23(2):252-259
An experiment was conducted to evaluate increasing or decreasing concentrations of dietary phytase on growth performance and processing yields of male broilers from 1 to 35 d of age. Treatments consisted of a positive control, a negative control (NC; less 0.14% Ca, 0.13% nonphytate P, and 0.03% Na), and 6 additional treatments based on the NC supplemented with phytase. Treatments 3 through 5 consisted of the NC diet supplemented with 500 phytase units (FTU)/kg of phytase in the starter phase that was either continued throughout the remainder of the study (treatment 3) or increased to 1,500 FTU/kg beginning in the finisher (treatment 4) or grower (treatment 5) phases. Treatment 6 had 1,500 FTU/kg of phytase throughout the study. Treatments 7 and 8 had 1,500 FTU/kg in the starter and decreased to 500 FTU/kg in the finisher or grower phases, respectively. At 35 d of age, broilers fed diets containing 1,500 FTU/kg of phytase had increased BW gain compared with birds fed diets formulated to contain 500 FTU/kg of phytase. Increasing phytase concentration between the starter and grower phases or decreasing phytase concentration between the grower and finisher phases negatively affected FCR from 1 to 35 d of age. Phytase supplementation did not affect weight and yield of carcass characteristics. Therefore, dietary phytase concentration should not be varied throughout production for optimum growth performance. 相似文献
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《The Journal of Applied Poultry Research》2008,17(2):191-201
A performance trial was conducted with broiler chicks to study the effect of phytase (PHY) supplementation in diets formulated with reduced AME, Ca, and P. The nutrient digestibility was determined during the 14- to 21-d and 28- to 35-d periods. The treatments consisted of 3 diets (NC1, NC2, NC3) differing in nutrient content and each diet with or without supplemental PHY (NC1, 0 or 500; NC2, 0 or 750; NC3, 0 or 1,000 U of PHY/kg feed) and 1 positive control diet (PC). Compared with the PC diet, negative control diets (NC) resulted in lower AME and apparent ileal amino acid digestibility for some amino acids. Phytase supplementation of the NC diets increased AME, apparent ileal amino acid digestibility, and apparent ileal crude protein digestibility. Phytase addition also increased mineral absorption in 21- and 35-d-old broilers fed NC diets. Reduced nutrient digestibility appears to be a factor in the weight gain and feed intake results. Reducing Ca and P content reduced feed intake in a stepwise fashion in the NC diets. Phytase increased feed intake and generally improved nutrient digestibility, which resulted in an increase in digestible nutrient intake. Averaged across NC diets, PHY improved body weight. Bone-breaking strength was the most consistent predictor of Ca and P reduction. All NC diets had significantly lower bone-breaking strength than the PC. Phytase supplementation of the NC diets gave bone-breaking strengths that were comparable to the PC. Diets with PHY had the highest bioeconomic index. 相似文献
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本文旨在研究小麦型日粮中添加木聚糖酶和植酸酶对肉鸡生长性能、回肠氨基酸表观消化率及矿物元素沉积的影响。试验选择1日龄科宝白羽肉仔鸡850只,随机分成5组,每组5个重复,每个重复34只鸡。一共有5种日粮,即低磷日粮、高磷日粮、低磷日粮+2000 IU/kg木聚糖酶、低磷日粮+500 FTU/kg植酸酶,低磷日粮+2000 IU/kg木聚糖酶+500 FTU/kg植酸酶,试验进行21 d。低磷组较高磷组显著降低了肉鸡的日增重、日采食量及趾骨灰分含量(P <0.05),与低磷组相比,木聚糖酶组显著降低了肉鸡的料比(P <0.05),而植酸酶组显著提高了肉鸡日增重、日采食量、趾骨灰分及饲料利用率(P <0.05)。复合酶组显著提高了15种氨基酸回肠表观消化率(P <0.05)。与低磷组相比,木聚糖酶组显著提高了表观消化能和可消化代谢能及回肠氮消化率(P <0.05),而植酸酶显著提高了氮沉积及回肠氮消化率(P <0.05),复合酶组显著提高了代谢能、可消化代谢能及氮沉积量(P <0.05)。植酸酶和复合酶组较高磷组显著提高了磷表观消化率和沉积量(P <0.05),但低磷组较高磷组显著提高了钙的表观消化率(P <0.05),植酸酶酶和复合酶组较其他3组显著提高了钾的沉积量(P <0.05)。结论 :小麦型日粮同时添加植酸酶和木聚糖酶对肉鸡生长和消化的影响要优于单独添加这两种酶。 相似文献