CN112626145A - 一种提高油脂利用率的莫能霉素生产方法 - Google Patents
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Abstract
本发明属于莫能菌素发酵技术领域,具体公开一种提高油脂利用率的莫能霉素生产方法,本发明通过在在发酵培养阶段结束以后的发酵液中添加1%的脂肪酶,于40℃保温反应30‑60min,搅拌速率100‑200rpm,再送去干燥制粒的操作,使脂肪酶对油脂进行部分水解以便于菌体充分代谢减少放罐油脂残余。降低补氨补碱浓度目的在于减少因局部碱浓度过高导致造化颗粒的形成,提升产品质量。
Description
技术领域
本发明属于莫能菌素发酵技术领域,尤其涉及一种提高油脂利用率的莫能霉素生产方法。
背景技术
莫能菌素以干扰球虫细胞内钠钾离子的正常交换,使大量的钠离子进入细胞内,造成钠离子在胞内浓度高,渗透压上升,更多的水分进入球虫内。 莫能菌素可以影响反刍动物瘤胃内能量代谢,改善瘤胃发酵,提高丙酸与乙酸的产出比例,降低挥发性脂肪酸,减少甲烷生成,提高蛋白质利用效率。在当前禁止在饲料中添加促生长类抗生素的情况下,莫能霉素作为抗球虫治疗作用仍可以继续使用。莫能霉素发酵代谢过程为油脂代谢,肉桂地链霉菌利用棕榈油或豆油作为主要碳源产生莫能菌素。目前已有结果显示发酵过程油耗较高,油控水平高,直接导致产品中油含量较高,影响产品质量和外观。
发明内容
本发明的目的是提供一种提高油脂利用率的莫能霉素生产方法,本发明中在油罐中加入合适剂量的脂肪酶,目的对油脂进行部分水解以便于菌体充分代谢,减少放罐油脂残余,降低补氨补碱浓度目的在于减少因局部碱浓度过高导致造化颗粒的形成,提升产品质量。
为达到上述目的,本发明采用的技术方案是:一种提高油脂利用率的莫能霉素生产方法,采用肉桂地链霉菌发酵生产莫能菌素的培养基,发酵工艺为三级,包括一级种子培养、二级种子培养和发酵培养,在发酵培养阶段结束以后的发酵液中添加1%的脂肪酶,于40℃保温反应30-60min,搅拌速率100-200rpm,再送去干燥制粒即可。
进一步的,一级种子培养采用培养基培养,一级种子培养基组成为:葡萄糖5g/L、糊精20g/L、豆粕粉15g/L、酵母粉3g/L、玉米蛋白粉5g,培养工艺周期为20-30h,培养温度30-34℃,转速200rpm,移种指标为pH 6.6-7.1,菌浓15-20%。
进一步的,二级种子培养采用培养基培养,二级种子培养基组成为:葡萄糖5g/L、碳酸钙2g/L、豆粕粉15g/L、酵母粉3g/L,培养工艺一级接二级接种量0.04%,培养周期10-20h,温度30-34℃,转速200rpm,通气量为0.5-1.0vvm, 移种指标:pH 6.6-7.1,菌浓15-20%。
进一步的,发酵培养采用发酵罐培养基,发酵罐培养基的组成为:葡萄糖20g/L、豆粕粉20g/L、油酸甲酯10g/L、豆油20g/L、硝酸钠2g/L、硫酸钠2g/L、磷酸氢二钾0.1g/L、氯化锰0.3g/L、抗坏血酸0.02g/L,轻质碳酸钙3g/L,泡敌0.2g/L,培养工艺:接种量8.0%,周期:300h,温度30-34℃,转速100rpm,通气量为0.5-1.0vvm,发酵培养过程通过添加补料培养基保持培养液PH为6.5、油控水平1.0-2.0%。
进一步的,所述干燥制粒包括将酶解过以后的发酵液打入预处理罐中,从罐口加入氢氧化钠,搅拌均匀,使溶液的pH值为9.0- 10,再加入碳酸钙搅拌35-50分钟,将料液全部打入预混罐,开启压力喷干塔,调节空压机压力O.35MPa,进料流量为300L/h,进风温度120-180℃,出风温度60-90℃,出料口温度50-60℃,收集产品。
进一步的,所述补料培养基为棕榈油、18-23%的氨水、10%的液碱、0.5%硫酸铵与0.05%磷酸混合液中的至少一种。
进一步的,所述碳酸钙的加量为碳酸钙的加量计算值±5%。
本发明具有的优点是:发酵过程油耗较高,油控水平高,直接导致产品中油含量较高,影响产品质量和外观,这一现象的主要原因为油脂为大分子脂肪酸,表观密度大,较为粘稠,分散不均匀;油脂难以利用,导致利用率不高,残留较高,本发明针对存在的技术问题在油罐中加入合适剂量的脂肪酶,对油脂进行部分水解以便于菌体充分代谢,其机理为由于产莫能霉素菌为龟裂链霉菌,他们以油脂作为碳源提供能量进行生长代谢,油脂进入菌体内会经过自身内部的脂肪酶等一系列酶分解成小分子脂肪酸,进而便于利用,油罐中添加脂肪酶目的在于提前将油脂预处理,将大分子链分解为小分子脂肪酸,便于龟裂链霉菌吸收利用从而产生更多的莫能菌素,减少放罐油脂残余,增加了原有培养基中碳源的充分利用,从而降低补氨补碱又可以减少因局部碱浓度过高导致造化颗粒的形成,提升产品质量。
具体实施方式
实施例
一种提高油脂利用率的莫能霉素生产方法,采用肉桂地链霉菌发酵生产莫能菌素的培养基,发酵工艺为三级,包括一级种子培养、二级种子培养和发酵培养,包括以下步骤:
1)一级种子培养采用培养基培养,一级种子培养基组成为:葡萄糖5g/L、糊精20g/L、豆粕粉15g/L、酵母粉3g/L、玉米蛋白粉5g,培养工艺周期为20-30h,培养温度30-34℃,转速200rpm,移种指标为pH 6.6-7.1,菌浓15-20%;
2)二级种子培养采用培养基培养,二级种子培养基组成为:葡萄糖5g/L、碳酸钙2g/L、豆粕粉15g/L、酵母粉3g/L,培养工艺一级接二级接种量0.04%,培养周期10-20h,温度30-34℃,转速200rpm,通气量为0.5-1.0vvm, 移种指标:pH 6.6-7.1,菌浓15-20%;
3)发酵培养采用发酵罐培养基,发酵罐培养基的组成为:葡萄糖20g/L、豆粕粉20g/L、油酸甲酯10g/L、豆油20g/L、硝酸钠2g/L、硫酸钠2g/L、磷酸氢二钾0.1g/L、氯化锰0.3g/L、抗坏血酸0.02g/L,轻质碳酸钙3g/L,泡敌0.2g/L,培养工艺:接种量8.0%,周期:300h,温度30-34℃,转速100rpm,通气量为0.5-1.0vvm,发酵培养过程通过添加补料培养基保持培养液PH为6.5、油控水平1.0-2.0%,补料培养基为棕榈油、18-23%的氨水、10%的液碱、0.5%硫酸铵与0.05%磷酸混合液中的至少一种;
4)提取制备成品:向步骤3)中发酵结束的发酵液中加入1%的脂肪酶,于40℃保温反应30-60min,搅拌速率100-200rpm,使酶解率在30-40%酶解结束,加热到70°C,打入预处理罐中,从罐口加入氢氧化钠,搅拌均匀,使溶液的pH值为9.0- 10,再加入碳酸钙搅拌35分钟,碳酸钙的加量为碳酸钙的加量计算值±5%,将料液全部打入预混罐,开启压力喷干塔,调节空压机压力O.35MPa,进料流量为300L/h,进风温度120-180℃,出风温度60-90℃,出料口温度50-60℃,收集产品。
对比实验
首先找到合适的水解工艺和脂肪酶种类,确定影响反应指标,评估影响因子重要性,将因子进行试验验证,确定最佳反应条件。通过单因子分析,分别确定水解工艺为添加酶量1%、水解温度40℃、反应时间1 h、pH和转速自然(自然状态下的PH值6.0、转速50-100rpm),水解率为游离脂肪酸与油脂的比值,具体结果见表1.
表1
本发明所用工艺,接种针刮斜面孢子后接种于一级摇瓶种子(40ml种子液),培养温度30-32℃,转速200rpm,培养20小时左右,移种指标pH7.0左右,菌浓不低于15%,镜检菌丝美兰深、粗壮,菌丝长且舒展;二级罐装液量为100L,接种量为0.04%,培养温度32℃,转速200rpm,通气量0.5-1.0vvm,当pH为6.6-7.0,菌浓不低于16%,培养周期16-24h;发酵罐装液量为500L,发酵接种量为8%(需二级种子40L),培养温度30-35℃,转速130rpm,通气量为0.4-1.0vvm根据溶氧不低于30%。发酵过程检测菌浓、pH、氨基氮、氨氮、总糖、残油、效价。发酵结束检测效价和残油。
采用实施例1在油罐中增加脂肪酶水解棕榈油工艺,添加量为1%,40℃反应30-60min,搅拌100-200rpm,保证水解率在30-40%。
对照组与实验组控制工艺、原料、种子等均一致,统计对照组和实验组放罐效价,放罐体积,平均油耗进行分析实验结果见表2.
实验结果表明实验组将油罐中棕榈油进行部分水解后补油量、油耗和放罐残油均有降低,每吨产品成本可降低300-500元左右。
Claims (7)
1.一种提高油脂利用率的莫能霉素生产方法,采用肉桂地链霉菌发酵生产莫能菌素的培养基,发酵工艺为三级,包括一级种子培养、二级种子培养和发酵培养,其特征在于:在发酵培养阶段结束以后的发酵液中添加1%的脂肪酶,于40℃保温反应30-60min,搅拌速率100-200rpm,再送去干燥制粒即可。
2.如权利要求1所述的提高油脂利用率的莫能霉素生产方法,其特征在于:一级种子培养采用培养基培养,一级种子培养基组成为:葡萄糖5g/L、糊精20g/L、豆粕粉15g/L、酵母粉3g/L、玉米蛋白粉5g,培养工艺周期为20-30h,培养温度30-34℃,转速200rpm,移种指标为pH 6.6-7.1,菌浓15-20%。
3.如权利要求1所述的提高油脂利用率的莫能霉素生产方法,其特征在于:二级种子培养采用培养基培养,二级种子培养基组成为:葡萄糖5g/L、碳酸钙2g/L、豆粕粉15g/L、酵母粉3g/L,培养工艺一级接二级接种量0.04%,培养周期10-20h,温度30-34℃,转速200rpm,通气量为0.5-1.0vvm, 移种指标:pH 6.6-7.1,菌浓15-20%。
4.如权利要求1所述的提高油脂利用率的莫能霉素生产方法,其特征在于:发酵培养采用发酵罐培养基,发酵罐培养基的组成为:葡萄糖20g/L、豆粕粉20g/L、油酸甲酯10g/L、豆油20g/L、硝酸钠2g/L、硫酸钠2g/L、磷酸氢二钾0.1g/L、氯化锰0.3g/L、抗坏血酸0.02g/L,轻质碳酸钙3g/L,泡敌0.2g/L,培养工艺:接种量8.0%,周期:300h,温度30-34℃,转速100rpm,通气量为0.5-1.0vvm,发酵培养过程通过添加补料培养基保持培养液PH为6.5、油控水平1.0-2.0%。
5.如权利要求1所述的提高油脂利用率的莫能霉素生产方法,其特征在于:所述干燥制粒包括将酶解过以后的发酵液打入预处理罐中,从罐口加入氢氧化钠,搅拌均匀,使溶液的pH值为9.0-10 ,再加入碳酸钙搅拌35-50分钟,将料液全部打入预混罐,开启压力喷干塔,调节空压机压力O.35MPa,进料流量为300L/h,进风温度120-180℃,出风温度60-90℃,出料口温度50-60℃,收集产品。
6.如权利要求4所述的提高油脂利用率的莫能霉素生产方法,其特征在于:所述补料培养基为棕榈油、18-23%的氨水、10%的液碱、0.5%硫酸铵与0.05%磷酸混合液中的至少一种。
7.如权利要求5所述的提高油脂利用率的莫能霉素生产方法,其特征在于:所述碳酸钙的加量为碳酸钙的加量计算值±5%。
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