CN109536268B - 一种混合真菌分段式固态发酵制备功能性油脂的方法 - Google Patents
一种混合真菌分段式固态发酵制备功能性油脂的方法 Download PDFInfo
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- CN109536268B CN109536268B CN201811569022.4A CN201811569022A CN109536268B CN 109536268 B CN109536268 B CN 109536268B CN 201811569022 A CN201811569022 A CN 201811569022A CN 109536268 B CN109536268 B CN 109536268B
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Abstract
本发明公开了一种混合真菌分段式固态发酵制备功能性油脂的方法,属于食用油加工领域。本发明利用大豆作为主要发酵底物,通过对不同功能营养的需求,搭配其他油籽,利用真菌分段式固态发酵,并结合超临界萃取分馏技术,以达到改善大豆油的回味和产生不同风味的目的,同时提高油脂的营养价值。本发明的真菌分段式固态发酵可以提高提油率,改善油脂脂肪酸组成,增加主要风味物质吡嗪类、醇类和酚类等物质的含量,其风味及色泽等感官功能更加符合消费者对创新产品的需求。发酵后的粕可以进一步加工,生产功能性多肽等副产物,实现可持续绿色发展。
Description
技术领域
本发明涉及一种混合真菌分段式固态发酵制备功能性油脂的方法,属于食用油加工领域。
背景技术
大豆油是世界上产量最多的植物油,传统的制备大豆油的方法有压榨法和浸提法,压榨法主要是利用螺旋榨油机进行压榨,压榨得到的大豆油能够保留其原有的风味和营养成分,但是其对原料的选料要求严格,且存在提油率偏低的问题,会大大的增加其生产成本。浸提法的原理是通过有机溶剂萃取大豆油,其过程需要经过高温脱臭等精炼过程,会破坏油中的影响成分,且对后期的精炼要求较高。现有技术中也公开了利用水酶法进行提油,尽管水酶法更加环保,原料利用率较高,但是其对设备要求较高,此外,由于油水分离困难,导致水酶法无法大规模发展与应用。
随着科学技术的发展,固态发酵以其独特的优点(如无“三废”排放)获得了人们的关注,固态发酵是指利用自然底物做碳源和能源,或是惰性底物做固体支持物,体系接近无水的发酵过程。现有技术中公开了利用固态发酵和水酶法或者冷榨结合的方式相结合的技术,对大豆或者芝麻进行发酵,能够提高提油率,但是,目前针对固态发酵的研究较少,尤其是对于不同菌株对此种工艺的影响研究也有待进一步探索完善,找到适当的菌株是影响提油效果与副产物品质的关键所在。
功能性油脂是一种对人体具有营养保健功能的油脂,不仅具有良好的感官功能(风味、外观),还具有比较高的营养功能。随着社会的发展和人们生活水平的提高,越来越多的人们追求健康生活,功能性油脂成为了越来越多的人的青睐。市场上为了满足消费者对功能性油脂的需求,衍生出一系列调和油和在一级油里面添加脂溶性维生素、酚类、甾醇类等物质。食用调和油是将两种或两种以上的植物油脂按比例调和配制的食用油,目前市场上调和油概念比较模糊,在调和油国家标准未颁布之前,未对调和油脂肪酸组成及有益伴随物有严格限制。很多商家为了赚取利润,并且假借调和油的概念大做文章,经常出现以低充高、以次充好等现象。
因此,急需寻找一种绿色、高效的生产功能性油脂的方法,以满足市场上对功能性油脂的需求。
发明内容
[技术问题]
本发明要解决的技术问题是提供一种可以满足消费者对创新油脂产品、油脂风味、色泽及功能性的追求的油脂及其制备方法,同时提高资源综合利用率。
为了实现上述目的,本发明提供的技术方案是综合真菌分段式固态发酵技术、超临界萃取分馏技术,制备针对不同人群所需的不同功能性油脂,具体地,本发明提供了一种混合真菌分段式固态发酵功能性油脂的方法,所述方法包括以下步骤:
(1)粉料准备:大豆破碎去皮后,轧坯,之后利用液压锥形塞膨化机进行膨化,膨化温度150~160℃,膨化时间1~3min,再冷却至25~35℃,得到大豆粉料;将其他油料种子剥壳或脱皮,粉碎至50~100目,粉碎后的粉料在115~121℃高压灭菌15~30min,冷却至25~35℃,得到其他油籽粉料;
(2)真菌发酵:将膨化后的大豆粉料与高压灭菌的其他油籽粉料配比为6~9:1~4(W/W)混合放入发酵罐,调节发酵罐中物料的水分含量为20~30wt%,葡萄糖添加量0.5~5wt%,NaCl添加量0.05~0.5wt%,调节pH为3.5~9,接种10~20wt%的真菌菌种,进行分段式发酵,发酵温度25~35℃,发酵时间为3~7d;
(3)超临界CO2萃取分馏:将发酵产物转入萃取釜,进行超临界CO2萃取,之后进行减压分馏,即可制备不同风味和色泽的营养强化的油脂,向油脂中充氮保藏;
其中,所述真菌为霉菌或酵母,所述霉菌包括米曲霉、黑曲霉和深黄被孢霉,所述酵母包括胶粘红酵母、白色假丝酵母和圆红冬孢酵母;所述分段式发酵为:当真菌为霉菌时,调节pH为3.5~9,溶氧量为总物料体积的30%~40%,先加入米曲菌进行发酵1~3d,之后补加0.1~1wt%的葡萄糖,调节pH为3.5~9,溶氧量为总物料体积的30%~40%,再添加黑曲霉和深黄被孢霉,培养2~6d;或,当真菌为酵母时,调节pH为5~9,溶氧量为总物料体积的25%~35%,先添加白色假丝酵母或白色假丝酵母和圆红冬孢酵母,发酵2~5d,之后补加1%~3%葡萄糖,调节pH为5~9,溶氧量为总物料体积的10%~20%再添加胶粘红酵母和圆红冬孢酵母,或胶粘红酵母,发酵1~4d。
在本发明的一种实施方式中,所述其他油料种子为亚麻籽、火麻籽、菜籽、芝麻、花生、玉米胚芽中的任一种或几种。
在本发明的一种实施方式中,所述米曲霉、黑曲霉和深黄被孢霉的质量比为1~5:1~5:1~5。
在本发明的一种实施方式中,所述胶粘红酵母、白色假丝酵母和圆红冬孢酵母的质量比为1~5:1~5:1~5。
在本发明的一种实施方式中,所述萃取压力30~40MPa,CO2流速为15~20L/h,萃取温度为40~50℃,萃取时间为1.5~2h。
在本发明的一种实施方式中,所述减压分馏中分为一级减压分馏和二级减压分馏,所述一级减压分馏的压力为10~20MPa,温度40~50℃;所述二级减压分馏的压力为8~10MPa,温度为35~45℃。
本发明取得的有益效果:
(1)本发明创新性的利用混合真菌发酵技术,通过真菌利用粉料生长代谢产酶等,将最主要产生豆腥味的2-戊基呋喃代谢或转化,并通过与其他油料种子如:亚麻籽、火麻籽、菜籽、花生等的协同作用来改善油脂风味,增加油脂中吡嗪类、醇类等风味物质的含量;本发明采用的胶粘红酵母、圆红冬孢酵母发酵制油,会赋予油脂明亮的红色,可以满足消费者对创新产品的渴望,更加推动油脂新产品开发;
(2)本发明方法通过混合真菌发酵从源头改善油脂中脂肪酸组成,获取高附加值脂肪酸(γ-亚麻酸、花生四烯酸、二十碳五烯酸、二十二碳六烯酸等),本发明制备得到的油脂中的不饱和脂肪酸含量最高可达84%,且本发明方法能够最大程度保留油脂中的营养因子,天然抗氧化剂的保留率可达90%;
(3)本发明利用液压锥形塞膨化机对大豆进行膨化,这种热处理方式对豆油营养物质几乎没有任何负面影响,特别是对蛋白质的质量无影响;并且还可以灭活抗营养因子,减少微生物污染,降低高压灭菌时的能耗;利用超临界萃取技术可以实现绿色环保生产,减少化学污染的产生;并且混合真菌发酵后的粕中,将含有大量功能性多肽,可以进一步深加工处理,提高产物附加值;
(4)本发明方法通过混合真菌分段式固态发酵,其出油率也大大增加,提油率最高可达95.0%,相比与未经发酵提油率增加40~60%,比单一菌种发酵的提油率可提高18~27%;
(5)本发明能够制备不同风味和色泽营养强化的油脂,能够满足消费者对创新油脂产品的需求,以及对油脂风味、色泽及功能性的追求,实现绿色环保生产,同时提高资源综合利用率。
附图说明
图1是一种混合真菌分段式固态发酵功能性油脂的方法示意图。
具体实施方式
出油率(%)=(原粉料中油脂含量-粉料粕中油脂含量)/原粉料中油脂含量×100%。
不饱和脂肪酸含量的计算方法:参考国标GB 5009.168-2016。
油脂中的有益伴随物的保留率:1-(索式提取得到油脂中有益伴随物的含量-发酵超临界提油有益伴随物的含量)/索式提取得到油脂中有益伴随物的含量×100%。
吡嗪类和醇类、酚类物质的相对含量的计算方法:主要风味物质含量/全部风味物质含量×100%。
霉菌活化:
米曲霉:将米曲霉接种于马铃薯培养液中,于180rpm 35℃条件下振荡培养2d;
黑曲霉:将黑曲霉接种于含3wt%葡萄糖、0.2wt%硫酸镁、0.2wt%磷酸二氢钾的种子培养基中,于180rpm 28℃培养2d;
深黄被孢霉:将深黄被孢霉接种于马铃薯培养液中,并加入0.05wt%葡萄糖,0.005wt%的磷酸二氢钾,0.015wt%硫酸镁,于180rpm 28℃培养2d。
酵母菌活化:
胶粘红酵母:将胶粘红酵母接种于含4wt%葡萄糖,0.3wt%酵母粉,0.2wt%硫酸铵,0.7wt%磷酸二氢钾,0.15wt%硫酸镁的培养基中,于180rpm 30℃条件下振荡培养6d;
白色假丝酵母:将白色假丝酵母接种于含2wt%葡萄糖,0.5wt%酵母粉,0.8wt%蛋白胨培养基中,于180rpm 30℃条件下振荡培养4d;
圆红冬孢酵母:将圆红冬孢酵母接种于含7wt%葡萄糖,0.01wt%硫酸铵,0.05wt%酵母粉,0.04wt%磷酸二氢钾,0.15wt%硫酸镁培养基中,于180rpm 30℃条件下振荡培养6d。
实施例1
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度150℃,膨化时间2min,冷却至35℃;将亚麻籽粉碎至100目,利用121℃高压灭菌15min,冷却至28℃;
(2)混合真菌发酵
将膨化后的大豆粉料和高压灭菌亚麻籽粉料按照8:2(W/W)混合,转入发酵罐中,调节水分含量为20wt%,添加葡萄糖1wt%,NaCl添加量0.05wt%,调节pH为5.0左右,溶解氧量为总物料体积的30%;将米曲霉加入发酵罐中,接种量为10%(W,下同),发酵温度为35℃,发酵时间为2d;2d后将发酵罐中的温度降低至28℃,补充0.5wt%的葡萄糖,将黑曲霉:深黄被孢霉按照1:1,总接种量为10%混合加入发酵罐中,pH自然,溶解氧量为总物料体积的40%,发酵温度为28℃,发酵5d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为15L/h,萃取温度为45℃,萃取时间为2h,之后进行减压分馏,一级减压分馏压力为15MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,出油率为91.7%,亚油酸、亚麻酸和花生四烯酸等不饱和脂肪酸含量为79.5%,油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留,保留率为92.0%,油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为30.8%,大豆油脂风味得到改善。
实施例2
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度160℃,膨化时间2min,冷却至30℃。将亚麻籽粉碎至100目,利用121℃高压灭菌15min,冷却至30℃;
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌亚麻籽粉料按照8:2(W/W)混合,转入发酵罐中,调节水分含量为20wt%,葡萄糖添加量为5wt%,NaCl添加量0.1%,调节pH为6.2,溶解氧量为总物料体积的30%,将白色假丝酵母:圆红冬孢酵母按照1:1(W/W),接种量为10%混合接种于发酵罐中,发酵温度为30℃,发酵3d;3d后将发酵温度降至28℃,补充2wt%葡萄糖,溶解氧量调节为总物料体积的15%,pH为调节为6.0,接种10%的粘红酵母,发酵温度28℃,发酵时间4d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为15L/h,萃取温度为40℃,萃取时间为1.5h。一级减压分馏压力为10MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到不同风味和色泽的功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为94.6%,脂肪酸组成变化不显著,油脂风味随着酵母发酵反应得到极大改善,油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为34.9%。且油脂色泽呈现微红,油脂色泽透亮,油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留,保留率为90.1%。
实施例3
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度160℃,膨化时间1min,冷却至35℃。将火麻籽破壳粉碎至100目,利用121℃高压灭菌15min,冷却至35℃;
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌火麻籽粉料按照9:1(W/W)混合,转入发酵罐中,调节水分含量为25wt%,添加葡萄糖5wt%,NaCl添加量0.05wt%,调节pH为5.0左右,溶解氧量为总物料体积的40%;将米曲霉加入发酵罐中,接种量为8%(W),发酵温度为35℃,发酵时间为2d;2d后将发酵罐中的温度降低至28℃,补充0.1wt%的葡萄糖,将黑曲霉:深黄被孢霉按照1:3,总接种量为8%混合加入发酵罐中,pH自然,溶解氧量为总物料体积的30%,发酵温度为28℃,发酵5d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力40MPa,CO2流速为20L/h,萃取温度为50℃,萃取时间为2h。一级减压分馏压力为20MPa,温度50℃;二级减压分馏压力为10MPa,温度为45℃,制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为89.3%,亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为84.3%,油脂中的有益伴随物维生素E、生育酚等天然抗氧化剂得到最大程度保留,保留率为90.6%,火麻籽油的风味以及发酵作用,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为48.7%,大豆油的“回味”完全消失。
实施例4
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度160℃,膨化时间1min,冷却至30℃。将亚麻籽粉碎至100目,利用121℃高压灭菌15min,冷却至30℃;
(2)混合真菌发酵
将膨化大豆粉料和高压灭菌火麻籽粉料按照9:1(W/W)混合,转入发酵罐中,调节水分含量为25wt%,葡萄糖添加量为4wt%,NaCl添加量0.2wt%,调节pH为6.2,溶解氧量为总物料体积的30%,将白色假丝酵母:圆红冬孢酵母按照1:1(W/W),接种量为8%混合接种于发酵罐中,发酵温度为30℃,发酵3d;3d后将发酵温度降至28℃,补充3wt%葡萄糖,溶解氧量调节为总物料体积的10%,pH为调节为6.0,接种10%的粘红酵母,发酵温度28℃,发酵时间4d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力35MPa,CO2流速为15L/h,萃取温度为45℃,萃取时间为2h。一级减压分馏压力为20MPa,温度40℃;二级减压分馏压力为8MPa,温度为40℃,制备得到不同风味和色泽的营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为22.1%,脂肪酸组成变化不显著,油脂风味随着酵母发酵反应得到极大改善,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为49.5%。且油脂色泽呈现深红,油脂色泽透亮,油脂中的有益伴随物维生素E、生育酚等酚类物质、植物甾醇等得到最大程度保留,保留率达到91.6%。
实施例5
(1)将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度150℃,膨化时间3min,冷却至35℃。将花生脱皮粉碎至150目,利用115℃高压灭菌30min,冷却至35℃;
(2)混合真菌发酵:
将膨化大豆粉料和高压灭菌花生粉料按照8:2(W/W)混合,转入发酵罐中,调节水分含量为30wt%,添加葡萄糖3wt%,NaCl添加量1wt%,调节pH为5.0左右,溶解氧量为总物料体积的40%;将米曲霉加入发酵罐中,接种量为8%(W),发酵温度为35℃,发酵时间为2d。2d后将发酵罐中的温度降低至28℃,补充0.1wt%的葡萄糖,将黑曲霉:深黄被孢霉按照1:3,总接种量为8%混合加入发酵罐中,pH自然,溶解氧量为总物料体积的30%,发酵温度为28℃,发酵5d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为20L/h,萃取温度为45℃,萃取时间为2h。一级减压分馏压力为15MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为94.9%,亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为80.5%,油脂中的有益伴随物维生素E、生育酚、三萜类、黄酮类等得到最大程度保留,保留率达到84.5%,花生油的风味以及发酵共同作用,可以改善豆油回味,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为20.9%。
实施例6
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度150℃,膨化时间3min,冷却至28℃。将花生脱皮粉碎至150目,利用115℃高压灭菌30min,冷却至30℃;
(2)混合真菌发酵:
将膨化大豆粉料和高压灭菌花生粉料按照8:2(W/W)混合,转入发酵罐中,调节水分含量为25wt%,葡萄糖添加量为5wt%,NaCl添加量0.2wt%,调节pH为6.2,溶解氧量为总物料体积的30%,将白色假丝酵母接种于发酵罐中,接种量为8%,发酵温度为30℃,发酵4d;4d后将发酵温度降至28℃,补充3wt%葡萄糖,溶解氧量调节为总物料体积的20%,pH为调节为5.8,圆红冬孢酵母:粘红酵母按1:1比例混合接种于发酵罐中,总接种量为10%,发酵温度28℃,发酵时间3d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为15L/h,萃取温度为40℃,萃取时间为1.5h。一级减压分馏压力为10MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到良好风味、色泽及营养强化功能性油脂通过充氮保藏。。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为95.0%,油脂中的有益伴随物维生素E、生育酚、三萜类、黄酮类等得到最大程度保留,保留率达到83.9%。花生油的风味以及发酵共同作用,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为18.7%,油脂呈现红色,且具有良好的透亮度。
实施例7
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度160℃,膨化时间2min,冷却至28℃。将菜籽脱皮粉碎至50目,利用121℃高压灭菌15min,冷却至28℃;
(2)混合真菌发酵:
将膨化大豆粉料和高压灭菌菜籽粉料按照3:2(W/W)混合,转入发酵罐中,调节水分含量为25wt%,添加葡萄糖1wt%,NaCl添加量0.05wt%,调节pH为5.0左右,溶解氧量为总物料体积的30%;将米曲霉加入发酵罐中,接种量为10%(W),发酵温度为35℃,发酵时间为2d;2d后将发酵罐中的温度降低至28℃,补充0.5wt%的葡萄糖,将黑曲霉:深黄被孢霉按照1:1,总接种量为10%混合加入发酵罐中,pH自然,溶解氧量为总物料体积的40%,发酵温度为28℃,发酵5d。
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为20L/h,萃取温度为45℃,萃取时间为2h。一级减压分馏压力为15MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为92.3%,亚油酸、亚麻酸和花生四烯酸不饱和脂肪酸含量为80.2%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等得到最大程度保留,保留率达到91.0%。大豆的回味和菜籽油的菜油味通过发酵得到有效改善,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为10.8%。
实施例8
(1)粉料准备
将大豆破碎、去皮、轧坯后,利用液压锥形塞膨化机对粉料膨化,膨化温度160℃,膨化时间2min,冷却至30℃。将菜籽脱皮粉碎至50目,利用121℃高压灭菌15min,冷却至30℃;
(2)混合真菌发酵:
将膨化大豆粉料和高压灭菌菜籽粉料按照3:2(W/W)混合,转入发酵罐中,调节水分含量为30wt%,葡萄糖添加量为5wt%,NaCl添加量0.2wt%,调节pH为6.2,溶解氧量为总物料体积的30%,将白色假丝酵母接种于发酵罐中,接种量为8%,发酵温度为30℃,发酵4d;4d后将发酵温度降至28℃,补充3wt%葡萄糖,溶解氧量调节为总物料体积的20%,pH为调节为5.8,圆红冬孢酵母:粘红酵母按1:1比例混合接种于发酵罐中,总接种量为10%,发酵温度28℃,发酵时间3d;
(3)超临界CO2萃取分馏
将发酵产物转入萃取釜中,萃取压力30MPa,CO2流速为15L/h,萃取温度为40℃,萃取时间为1.5h。一级减压分馏压力为10MPa,温度40℃;二级减压分馏压力为8MPa,温度为35℃,制备得到良好风味及营养强化功能性油脂通过充氮保藏。
通过该混合真菌分段式固态发酵方法制备油脂,提油率为96.1%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等得到最大程度保留,保留率达到84.1%,通过酵母发酵可以有效改善豆油豆腥味及菜籽油的菜油味,发酵油脂中主要风味物质吡嗪类和醇类、酚类等物质相对含量为12.3%,发酵后油脂呈现暗红色。
对比例1
单独利用米曲霉进行发酵,发酵过程为:将米曲霉加入发酵罐中,总的接种量为20%(W),分段发酵,pH5.0,发酵温度为35℃,通风发酵,总的发酵时间为7d,其余步骤和条件与实施例1发酵条件相同,此时的出油率为74.2%,不饱和脂肪酸含量为64.5%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为76.3%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为7.3%。
同样的,单独利用黑曲霉进行发酵,发酵过程为:将黑曲霉加入发酵罐中,总的接种量为20%(W),分段发酵,pH自然,发酵温度为28℃,通风发酵,总的发酵时间为7d,其余步骤和条件与实施例1发酵条件相同,此时的出油率为73.3%,不饱和脂肪酸含量为62.8%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为59.6%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为6.4%。
同样的,单独利用深黄被孢霉进行发酵,发酵过程为:将深黄被孢霉加入发酵罐中,总的接种量为20%(W),pH自然,发酵温度为28℃,通风发酵,总的发酵时间为7d,其余步骤和条件与实施例1发酵条件相同,此时的出油率为78.6%,不饱和脂肪酸含量为68.1%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为70.2%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为8.1%。
利用单独的米曲霉、黑曲霉或深黄被孢霉进行发酵,在发酵3~4d后菌种活力大幅度下降,发酵底物得不到充分利用。如果不及时补充碳源和菌种,会降低发酵提油的效率。通过多菌种混合分段式发酵,能够显著的提高出油率,提高了约16%~25%;此外,本发明的混合真菌分段发酵能够充分发挥各菌种的对脂肪酸的协同转化能力,提高油脂中不饱和脂肪酸的含量,提高了约17%~25%;利用多菌种分段式固态发酵结合超临界二氧化碳萃取的方法,可以提高油脂中有益伴随物的保留率,可增加17~25%;此外,本发明方法能够显著的增加油脂中吡嗪类、酚类、醇类及酸类等主要风味物质的含量,明显的改善大豆油产品中的回味。
对比例2
单独利用胶粘红酵母进行发酵,发酵过程为:将胶粘红酵母加入发酵罐中,总的接种量为20%(W),pH6.2,发酵温度为30℃,通风发酵,发酵总时间为7d,其余步骤和条件与实例2发酵条件相同,此时的出油率为78.5%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为67.9%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为14.5%。
单独利用白色假丝酵母进行发酵,发酵过程为:将白色假丝酵母加入发酵罐中,总的接种量为20%(W),pH6.0,发酵温度为30℃,通风发酵,发酵总时间为7d,其余步骤和条件与实施例2发酵条件相同,此时的出油率为80.2%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为80.4%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为13.9%。
单独利用圆红冬孢酵母进行发酵,发酵过程为:将圆红冬孢酵母加入发酵罐中,总的接种量为20%(W),pH6.0,发酵温度为30℃,通风发酵,发酵总时间为7d,其余步骤和条件与实施例2发酵条件相同,此时的出油率为77.9%,油脂中的有益伴随物维生素E、生育酚、植物甾醇等保留率为73.8%,主要风味物质吡嗪类、醇类及酚类等物质相对含量为13.4%。
可见,利用单一酵母发酵无法达到完全发酵,菌种在发酵一段时间会失去活力。相比而言,本发明方法通过多菌种混合分段式发酵结合超临界二氧化碳萃取的方法,能够保持菌种的持续活力,能够明显地提高提油率,提高了约18~20%,并能够在一定程度上提高维生素E、甾醇、三萜及黄酮类等有益伴随物的含量,可增加22~33%;且油脂中主要风味物质吡嗪类、醇类、酚类及酸类的物质含量大大增加。
对比例3
先添加黑曲霉:深黄被孢霉进行固态发酵,再添加米曲霉进行发酵,其余步骤和条件与实施例1发酵条件相同,此时的提油率为86.4%,不饱和脂肪酸含量为72.3%,主要风味物质吡嗪类和醇类、酚类等的相对含量为16.3%。改变三种霉菌的添加顺序,则导致其提油率下降,尤其的,主要风味物质的含量明显降低,仅有实施例1中的50%左右;此外,改变发酵顺序后,在接种米曲霉时,需要重新调整发酵pH值,增加人力成本。
对比例4
先添加粘红酵母:圆红冬孢酵母进行固态发酵,再添加白色假丝酵母进行发酵,其余步骤和条件与实施例2发酵条件相同,此时的提油率为87.3%,不饱和脂肪酸含量为70.6%,主要风味物质吡嗪类和醇类、酚类等的相对含量为18.6%;此外,改变发酵顺序后,发酵得到油脂颜色没有实施例2制备得到的油脂颜色鲜亮。
对比例5
若大豆:其他油籽为2:3时,利用实施例1和2的方法进行固态发酵,此时提油率分别为86.3%和87.5%。其他油籽用量增多,会在一定程度上增加原料成本。此外,亚麻油、火麻油及花生油等油脂自身具有很强的风味,较大的含量时,无需发酵即能够很好掩盖大豆油脂的回味,若再用发酵提油又会进一步增加成本;而且亚麻油、火麻油及菜籽油等本身就含有大量不饱和脂肪酸,且含有产生风味的吡嗪类等主要物质,若增大其比例,会增加油脂中2-戊基吡啶的含量,容易产生不愉快的气味。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (8)
1.一种混合真菌分段式固态发酵制备功能性油脂的方法,其特征在于,所述方法包括以下步骤:
(1)粉料准备:大豆破碎去皮后,轧坯,之后利用液压锥形塞膨化机进行膨化,膨化温度150~160℃,膨化时间1~3min,再冷却至25~35℃,得到大豆粉料;将其他油料种子剥壳或脱皮,粉碎至50~100目,粉碎后的粉料在115~121℃高压灭菌15~30min,冷却至25~35℃,得到其他油籽粉料;
(2)真菌发酵:将膨化后的大豆粉料与高压灭菌的其他油籽粉料按照质量配比为6~9:1~4混合放入发酵罐,调节发酵罐中物料的水分含量为20~30wt%,葡萄糖添加量0.5~5wt%,NaCl添加量0.05~0.5wt%,调节pH为3.5~9,接种10~20wt%的真菌菌种,进行分段式发酵,发酵温度25~35℃,发酵时间为3~7d;
(3)超临界CO2萃取分馏:将发酵产物转入萃取釜,进行超临界CO2萃取,之后进行减压分馏,即可制备不同风味和色泽的营养强化的油脂,向油脂中充氮保藏;
其中,所述真菌为霉菌或酵母,所述霉菌包括米曲霉、黑曲霉和深黄被孢霉,所述酵母包括胶粘红酵母、白色假丝酵母和圆红冬孢酵母;所述分段式发酵为:当真菌为霉菌时,调节pH为3.5~9,溶氧量为总物料体积的30%~40%,先加入米曲菌进行发酵1~3d,之后补加0.1~1wt%的葡萄糖,调节pH为3.5~9,溶氧量为总物料体积的30%~40%,再添加黑曲霉和深黄被孢霉,培养2~6d,所述米曲霉、黑曲霉和深黄被孢霉的质量比为1~5:1~5:1~5;或,当真菌为酵母时,调节pH为5~9,溶氧量为总物料体积的25%~35%,先添加白色假丝酵母,或白色假丝酵母和圆红冬孢酵母,发酵2~5d,之后补加1%~3wt%葡萄糖,调节pH为5~9,溶氧量为总物料体积的10%~20%再添加胶粘红酵母和圆红冬孢酵母,或胶粘红酵母,发酵1~4d,所述胶粘红酵母、白色假丝酵母和圆红冬孢酵母的质量比为1~5:1~5:1~5。
2.根据权利要求1所述的一种混合真菌分段式固态发酵制备功能性油脂的方法,其特征在于,所述其他油料种子为亚麻籽、火麻籽、菜籽、芝麻、花生、玉米胚芽中的任一种或几种。
3.根据权利要求1或2所述的一种混合真菌分段式固态发酵制备功能性油脂的方法,所述萃取压力30~40MPa,CO2流速为15~20L/h,萃取温度为40~50℃,萃取时间为1.5~2h。
4.根据权利要求1或2所述的一种混合真菌分段式固态发酵制备功能性油脂的方法,其特征在于,所述减压分馏中分为一级减压分馏和二级减压分馏,所述一级减压分馏的压力为10~20MPa,温度40~50℃;所述二级减压分馏的压力为8~10MPa,温度为35~45℃。
5.根据权利要求3所述的一种混合真菌分段式固态发酵制备功能性油脂的方法,其特征在于,所述减压分馏中分为一级减压分馏和二级减压分馏,所述一级减压分馏的压力为10~20MPa,温度40~50℃;所述二级减压分馏的压力为8~10MPa,温度为35~45℃。
6.权利要求1~5任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法制备得到的油脂。
7.包含权利要求6所述的油脂的食品或食用油。
8.权利要求1~5任一所述的一种混合真菌分段式固态发酵制备功能性油脂的方法在食品加工领域的应用。
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