豆粕生物肽的生产工艺及其对肉鸡生产性能和血液生化指标的影响研究
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摘要
本文通过菌种筛选、摇瓶发酵条件优化、50L发酵罐扩大培养和发酵产物性质分析，系统地研究了利用微生物发酵生产豆粕生物肽的工艺，并对其在肉鸡日粮中的应用效果进行了研究。选用艾维因肉仔鸡320只，随机分为5组，每组4个重复，每个重复16只。第一组为对照组，饲喂基础日粮；其余4组分别添加0.1％、0.5％、1.0％、1.5％的豆粕生物肽。研究在饲粮中添加豆粕生物肽对肉仔鸡生产性能、血液常规、免疫以及对盲肠微生物等项指标的影响。
豆粕生物肽生产工艺研究确定：(1)细菌fj625作为本试验出发菌株；(2)摇瓶优化的适宜发酵条件为：豆粕15％，葡萄糖1％～4％，氯化镁0.02％，磷酸氢二钾0.01％，初始pH为6～7，豆粕粒度＜40目；(3)发酵罐扩大培养培养基组成为：豆粕15％，葡萄糖1.5％，氯化镁0.02％，磷酸氢二钾0.01 ％；发酵罐装液量为发酵罐容积的50％；消泡剂用量为有机硅0.04％和PDA0.25％；自然pH；发酵罐接种量为发酵体积的5％；(4)发酵过程中控制发酵温度为30℃；通过分阶段控制搅拌速度和通气量控制溶氧；维持罐压0.5Mpa。
豆粕生物肽常规营养成分测定结果为蛋白质49.27％，能量16.64MJ／kg，水分11.48％，粗脂肪3.02％，粗纤维11.84％，粗灰分8.6％，钙0.38％，磷0.87％，无氮浸出物15.79％。本试验产物豆粕生物肽分子量均小于8000Da，蛋白质降解为多肽的得率为42.01％，中性蛋白酶酶活为1538±35.26U／g。
动物试验结果表明：(1)肉鸡日粮中添加豆粕生物肽对前期、后期和全期的日增重均随添加量的增加逐渐增加，至添加量为1.5％时，与对照组相比差异极显著(p＜0.01)。从全期来看，日增重分别比对照组增加了3.03％、3.52％、4.60％、7.40％。(2)对平均日耗料的影响，从全期来看，各试验组均高于对照组，分别比对照组增加了4.75％、4.54％、3.33％、3.06％。其中第2、3组与对照组相比差异显著(P＜0.05)，各试验组随着日粮中豆粕发酵肽粉添加量的增加，平均日耗料呈下降的趋势。(3)对料肉比的影响，试验前期，日粮中添加量为1.5％时，料肉比降低了7.36％，显著低于对照组(P＜0.05)。其它各试验组与对照组差异不显著。(4)试验前期，日粮中添加豆粕生物肽1.0％、1.5％每公斤增重分别比对照组节约饲料成本0.09 yuan／kg和0.15yuan／kg。后期，添加1.5％节约饲料成本0.05 yuan／kg。(5)日粮中添加豆粕生物肽，对21日龄肉仔鸡血清蛋白无显著影响(P＞0.05)，血清尿素氮各试验组与对照组差异不显著(P＞0.05)。42日龄肉仔鸡血清测定结果表明，各试验组血清总蛋白含量均高于对照组，其中第4组与对照组差异显著(P＜0.05)。血清球蛋白各试验组间差异不显著(P＞0.05)。各试验组血清白蛋白含量均高于对照组，其中第3组与对照组差异显著(P＜0.05)，第4组和第5组与对照组差异极显著(P＜0.01)。血清尿素氮各组间差异不显著。(6)试验日粮中添加豆粕发酵肽粉对21日龄肉仔鸡免疫器官(脾脏、胸腺和法氏囊)指数无显著影响。42日龄肉仔鸡脾脏指数，第3组与对照组差异极显著(P＜0.01)，第4组与对照组差异显著(P＜0.05)。法氏囊指数随豆粕生物肽添加
量的增加逐渐增加，与21日龄的趋势相同。其中试验3组和4组与对照组之间差异
显著(P<0.05):第5组与对照组之间差异极显著(P<0.01)。‘7)肉鸡21日龄与对
照组相比随着添加量的增加，盲肠中大肠杆菌数量逐渐减少。当添加量为1 .0%时，
大肠杆菌降低数量与对照组相比差异显著(P<0.05)，添加量为1.5%时，与对照组差
异极显著(P<0.01)。42日龄，盲肠大肠杆菌计数，试验组均低于对照组，随添加量
的增加，盲肠中大肠杆菌数量有下降趋势，但各试验组与对照组相比均未达到显著水
平。(幻添加大豆降解肤粉对胸肌嫩度有增加的作用，21日龄屠宰的肉仔鸡胸肌嫩
度除第2组外，其余各组均比对照组增加，但未达显著水平(P<0.05)。42日龄屠宰
的肉仔鸡胸肌嫩度第4组与对照组差异极显著(P<0.01)，第5组与对照组差异显著
(P<().05)。
In this test, the production method of using bacterium fermenting soybean meal through selecting strain, optimizing the condition in the spinner flask, culturing in the fermenter and analysing the character of the ferment production was studied. And the effect of the production on the broilers was studied. 320 1-day-old Avain broilers were randomly allotted into five groups with 4 replicates of 15 birds each replicate. Group 1 was the basic group, fed basic diet, the other four groups were fed with basic diets adding 0.1%, 0.5%, 1.0%, 1.5% the biologic peptide of soybean meal respectively, studying the effect on the production performance, serum, immunity and the caecum microorganism of broilers.
The processing technology of the biologic peptide of soybean meal was determined as follows: (1) bacterium fj625 was the fermented strain; (2) the optimum conditions in the spinner flask were: soybean meal 15%, glucose 1%~4%, MgCl2 0.02%, K2HPO3 0.01%, pH 6-7, granularity of the soybean meal < 40 holes; (3) the fermentation medium of the fermentater contained: soybean meal 15%, glucose 1.5%, MgCl2 0.02%, K2HPO3 0.01%; the fermentated volume occupied by 50% of fermenter cubage; natural pH; inoculation volume 5%. (4) during the period of fermentation, the temperature was 30C, controlled the density of oxygen by controlling the agitation speed and ventilate; maintained the pressure at 0.5Mpa.
The nutrition level of the biologic peptide of soybean meal were: CP 49.27%, TE 16.64MJ/kg, EE 3.02%, CF 11.84%, Ash 8.6%, Ca 0.38%, TP 0.87%, NFE 15.79%. The molecular weight of the biologic peptide of soybean meal were all smaller 8000Da, the ratio of the peptides from the soybean protein was 42.01%, neutral proteinase activity was 1538?5.26U/g.
The result of animal experiment showed that: (1) the average daily gain of the experiment groups increased with the increasing of adding level of the biologic peptide of soybean meal. It was significant difference between the group 5 (adding 1.5%) and the group l(p<0.01). In the whole period (0~6w), comparing to group 1, the average daily gain of the group 2, 3, 4, 5 were increased by 3.03%, 3.52%, 4.60%, 7.40% (2) In the whole period (0~6w), comparing to group 1, the average daily feed intake of the group 2, 3, 4, 5 were increased by 4.75%, 4.54% 3.33% 3.06%. The group 4 and group5 could increase
the average daily feed intake significantly (p<0.05). With the increasing level of the biologic peptide of soybean meal, the average daily feed intake was in the downward. (3) In the prephase period (0-3w), comparing to control group, adding 1.5% biologic peptide of soybean meal to broiler diets decreased F/G by 7.36%(p<0.05). (4) In the prephase period (0~3w), the diets containing 1.0%, 1.5% biologic peptide of soybean meal saved the feed cost 0.09 yuan/kg and 0.15 yuan/kg respectively. In the anaphase period (4~6w), the diets containing 1.5% biologic peptide of soybean meal saved the feed cost 0.05 yuan/kg. (5) It was no significant difference of the serum protein and serum urea nitrogen of the 21-day broilers (p>0.05). Comparing to the control group, the TP of the 42-day broiler's serum of the experiment groups were increased and there was significant difference between group 4 and the control group (p<0.05). There were no significant differences of the globin among the experiment groups (p>0.05). The con
tent of the albumin of the experiment groups adding the biologic peptide of soybean meal were higher than control group. It was significant difference between the group 3 and the control group (p<0.05). Group 4 and Group 5 could increase the albumin significantly (p<0.01). The serum urea nitrogen was not significant changed (p>.05). (6) There were no significant differences of the immunity organs index by adding the biologic peptide of soybean meal. Increasing the spleen index of the 42-day broiler. It was significant difference between the group 3 and the group 1 (p<0.01) and it was significant difference between the group 4 and the groupl (p<0.05). The trend was same as the 21-day's. There w
豆粕生物肽生产工艺研究确定：(1)细菌fj625作为本试验出发菌株；(2)摇瓶优化的适宜发酵条件为：豆粕15％，葡萄糖1％～4％，氯化镁0.02％，磷酸氢二钾0.01％，初始pH为6～7，豆粕粒度＜40目；(3)发酵罐扩大培养培养基组成为：豆粕15％，葡萄糖1.5％，氯化镁0.02％，磷酸氢二钾0.01 ％；发酵罐装液量为发酵罐容积的50％；消泡剂用量为有机硅0.04％和PDA0.25％；自然pH；发酵罐接种量为发酵体积的5％；(4)发酵过程中控制发酵温度为30℃；通过分阶段控制搅拌速度和通气量控制溶氧；维持罐压0.5Mpa。
豆粕生物肽常规营养成分测定结果为蛋白质49.27％，能量16.64MJ／kg，水分11.48％，粗脂肪3.02％，粗纤维11.84％，粗灰分8.6％，钙0.38％，磷0.87％，无氮浸出物15.79％。本试验产物豆粕生物肽分子量均小于8000Da，蛋白质降解为多肽的得率为42.01％，中性蛋白酶酶活为1538±35.26U／g。
动物试验结果表明：(1)肉鸡日粮中添加豆粕生物肽对前期、后期和全期的日增重均随添加量的增加逐渐增加，至添加量为1.5％时，与对照组相比差异极显著(p＜0.01)。从全期来看，日增重分别比对照组增加了3.03％、3.52％、4.60％、7.40％。(2)对平均日耗料的影响，从全期来看，各试验组均高于对照组，分别比对照组增加了4.75％、4.54％、3.33％、3.06％。其中第2、3组与对照组相比差异显著(P＜0.05)，各试验组随着日粮中豆粕发酵肽粉添加量的增加，平均日耗料呈下降的趋势。(3)对料肉比的影响，试验前期，日粮中添加量为1.5％时，料肉比降低了7.36％，显著低于对照组(P＜0.05)。其它各试验组与对照组差异不显著。(4)试验前期，日粮中添加豆粕生物肽1.0％、1.5％每公斤增重分别比对照组节约饲料成本0.09 yuan／kg和0.15yuan／kg。后期，添加1.5％节约饲料成本0.05 yuan／kg。(5)日粮中添加豆粕生物肽，对21日龄肉仔鸡血清蛋白无显著影响(P＞0.05)，血清尿素氮各试验组与对照组差异不显著(P＞0.05)。42日龄肉仔鸡血清测定结果表明，各试验组血清总蛋白含量均高于对照组，其中第4组与对照组差异显著(P＜0.05)。血清球蛋白各试验组间差异不显著(P＞0.05)。各试验组血清白蛋白含量均高于对照组，其中第3组与对照组差异显著(P＜0.05)，第4组和第5组与对照组差异极显著(P＜0.01)。血清尿素氮各组间差异不显著。(6)试验日粮中添加豆粕发酵肽粉对21日龄肉仔鸡免疫器官(脾脏、胸腺和法氏囊)指数无显著影响。42日龄肉仔鸡脾脏指数，第3组与对照组差异极显著(P＜0.01)，第4组与对照组差异显著(P＜0.05)。法氏囊指数随豆粕生物肽添加
量的增加逐渐增加，与21日龄的趋势相同。其中试验3组和4组与对照组之间差异
显著(P<0.05):第5组与对照组之间差异极显著(P<0.01)。‘7)肉鸡21日龄与对
照组相比随着添加量的增加，盲肠中大肠杆菌数量逐渐减少。当添加量为1 .0%时，
大肠杆菌降低数量与对照组相比差异显著(P<0.05)，添加量为1.5%时，与对照组差
异极显著(P<0.01)。42日龄，盲肠大肠杆菌计数，试验组均低于对照组，随添加量
的增加，盲肠中大肠杆菌数量有下降趋势，但各试验组与对照组相比均未达到显著水
平。(幻添加大豆降解肤粉对胸肌嫩度有增加的作用，21日龄屠宰的肉仔鸡胸肌嫩
度除第2组外，其余各组均比对照组增加，但未达显著水平(P<0.05)。42日龄屠宰
的肉仔鸡胸肌嫩度第4组与对照组差异极显著(P<0.01)，第5组与对照组差异显著
(P<().05)。
In this test, the production method of using bacterium fermenting soybean meal through selecting strain, optimizing the condition in the spinner flask, culturing in the fermenter and analysing the character of the ferment production was studied. And the effect of the production on the broilers was studied. 320 1-day-old Avain broilers were randomly allotted into five groups with 4 replicates of 15 birds each replicate. Group 1 was the basic group, fed basic diet, the other four groups were fed with basic diets adding 0.1%, 0.5%, 1.0%, 1.5% the biologic peptide of soybean meal respectively, studying the effect on the production performance, serum, immunity and the caecum microorganism of broilers.
The processing technology of the biologic peptide of soybean meal was determined as follows: (1) bacterium fj625 was the fermented strain; (2) the optimum conditions in the spinner flask were: soybean meal 15%, glucose 1%~4%, MgCl2 0.02%, K2HPO3 0.01%, pH 6-7, granularity of the soybean meal < 40 holes; (3) the fermentation medium of the fermentater contained: soybean meal 15%, glucose 1.5%, MgCl2 0.02%, K2HPO3 0.01%; the fermentated volume occupied by 50% of fermenter cubage; natural pH; inoculation volume 5%. (4) during the period of fermentation, the temperature was 30C, controlled the density of oxygen by controlling the agitation speed and ventilate; maintained the pressure at 0.5Mpa.
The nutrition level of the biologic peptide of soybean meal were: CP 49.27%, TE 16.64MJ/kg, EE 3.02%, CF 11.84%, Ash 8.6%, Ca 0.38%, TP 0.87%, NFE 15.79%. The molecular weight of the biologic peptide of soybean meal were all smaller 8000Da, the ratio of the peptides from the soybean protein was 42.01%, neutral proteinase activity was 1538?5.26U/g.
The result of animal experiment showed that: (1) the average daily gain of the experiment groups increased with the increasing of adding level of the biologic peptide of soybean meal. It was significant difference between the group 5 (adding 1.5%) and the group l(p<0.01). In the whole period (0~6w), comparing to group 1, the average daily gain of the group 2, 3, 4, 5 were increased by 3.03%, 3.52%, 4.60%, 7.40% (2) In the whole period (0~6w), comparing to group 1, the average daily feed intake of the group 2, 3, 4, 5 were increased by 4.75%, 4.54% 3.33% 3.06%. The group 4 and group5 could increase
the average daily feed intake significantly (p<0.05). With the increasing level of the biologic peptide of soybean meal, the average daily feed intake was in the downward. (3) In the prephase period (0-3w), comparing to control group, adding 1.5% biologic peptide of soybean meal to broiler diets decreased F/G by 7.36%(p<0.05). (4) In the prephase period (0~3w), the diets containing 1.0%, 1.5% biologic peptide of soybean meal saved the feed cost 0.09 yuan/kg and 0.15 yuan/kg respectively. In the anaphase period (4~6w), the diets containing 1.5% biologic peptide of soybean meal saved the feed cost 0.05 yuan/kg. (5) It was no significant difference of the serum protein and serum urea nitrogen of the 21-day broilers (p>0.05). Comparing to the control group, the TP of the 42-day broiler's serum of the experiment groups were increased and there was significant difference between group 4 and the control group (p<0.05). There were no significant differences of the globin among the experiment groups (p>0.05). The con
tent of the albumin of the experiment groups adding the biologic peptide of soybean meal were higher than control group. It was significant difference between the group 3 and the control group (p<0.05). Group 4 and Group 5 could increase the albumin significantly (p<0.01). The serum urea nitrogen was not significant changed (p>.05). (6) There were no significant differences of the immunity organs index by adding the biologic peptide of soybean meal. Increasing the spleen index of the 42-day broiler. It was significant difference between the group 3 and the group 1 (p<0.01) and it was significant difference between the group 4 and the groupl (p<0.05). The trend was same as the 21-day's. There w
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