CN111500556A - 一种用于提高和稳定啤酒发酵度的复合酶制剂及其应用 - Google Patents
一种用于提高和稳定啤酒发酵度的复合酶制剂及其应用 Download PDFInfo
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- CN111500556A CN111500556A CN202010298232.5A CN202010298232A CN111500556A CN 111500556 A CN111500556 A CN 111500556A CN 202010298232 A CN202010298232 A CN 202010298232A CN 111500556 A CN111500556 A CN 111500556A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C5/00—Other raw materials for the preparation of beer
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- C12C5/006—Beta-glucanase or functionally equivalent enzymes
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- C12C7/00—Preparation of wort
- C12C7/04—Preparation or treatment of the mash
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- C12N9/2428—Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
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- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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- C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
- C12N9/2457—Pullulanase (3.2.1.41)
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
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- C12Y302/01041—Pullulanase (3.2.1.41)
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Abstract
本发明提供了一种用于提高和稳定啤酒发酵度的复合酶制剂,包括复合酶及辅料,所述复合酶包括如下单酶组分:中温α‑淀粉酶、葡糖淀粉酶、普鲁兰酶、中性蛋白酶、氨肽酶、半纤维素酶及β‑葡聚糖酶。另外,本发明还提供了将该复合酶制剂用于啤酒发酵的方法。该发明通过将不同淀粉酶和蛋白酶与葡聚糖酶和半纤维素酶进行复配后,有效地水解啤酒原料麦芽和配料中的淀粉类和非淀粉类多糖,增加麦汁中的可发酵性糖,达到稳定麦汁中的可发酵性糖和α‑N的含量的目的,进而低成本地提高麦汁的发酵度和稳定性,解决不同批次麦芽啤酒发酵的不稳定的问题。
Description
技术领域
本发明属于生物酶制剂技术领域,具体涉及一种用于提高和稳定啤酒发酵度的复合酶制剂及其应用。
背景技术
我国是世界上规模最大的饮料酒生产和消费国,其中啤酒作为饮料酒的重要组成部分,在国内市场规模很庞大。啤酒是以麦芽为主要原料,其他谷物为辅料,经过原辅料的纯净化处理,糖化过程中麦芽汁的准备、过滤及煮沸(该过程中需添加酒花或酒花制品),进而在发酵阶段添加不同啤酒酵母进行发酵后,酿制所得的低酒精度、起泡、含二氧化碳的酒精饮料。影响啤酒的质量的不稳定因素有很多,这其中啤酒发酵度指标的控制意义重大,它不仅可以反映产品相应批次原料质量的好坏,还可以对其生产过程中各项参数控制的稳定性、优劣性进行间接地判定;啤酒发酵度控制的稳定与否,直接影响到产成品的部分描述性指标,如啤酒的口感、风味、味道。
而啤酒的发酵度指标较为敏感,影响啤酒真正发酵度的因素有很多,主要包括:麦芽和辅料的选用及处理、糖化过程中工艺条件的设定及控制、较适宜的发酵工艺、对酵母菌株的选育,以及与麦汁接触的通氧量、酵母菌株的接种量、成品啤酒与氧含量之间的关系等发酵过程,这其中大部分的因素在啤酒厂的实际操作中都可以控制,但是由于大麦的生长受环境影响,制麦技术不同,在实际生产中发现,不同批次的麦芽在相同条件下模拟糖化,极限发酵度有较大的不同,有报道其中偏差高达6%,而事实上,1%的差异就会直接影响产品的品质。
为此不同批次麦芽进仓时都需要检测极限发酵度来进一步调整后续工艺,福建燕京惠泉啤酒公司在专利CN 106753946A中通过抽样测定麦芽极限发酵度,根据批次不同微调麦芽的配比来解决,然而这种方法在麦芽批次比较多的时候,耗时耗力,而且无法解决同一批次麦芽发酵度低的问题。此外,随着人们生活水平的提高,对于健康生活也日益关注,无糖食品也越来越受到消费者青睐,在啤酒行业中,无糖低卡里路啤酒成为一个热门研究热点,这其中的主要途径就是通过提高麦汁发酵度实现低糖或无糖。生物酶制剂目前广泛应用于食品加工行业,在啤酒行业应用历史也很丰富,但是在目前申请公开的专利中还未发现用生物酶法稳定和提高啤酒发酵度的方法。
发明内容
本发明的目的是克服现有啤酒糖化发酵过程中可发酵性糖和α-N的含量不稳定,影响啤酒发酵度进而影响啤酒品质的问题。
为此,本发明提供了一种用于提高和稳定啤酒发酵度的复合酶制剂,包括复合酶及辅料,所述复合酶包括如下单酶组分:中温α-淀粉酶、葡糖淀粉酶、普鲁兰酶、中性蛋白酶、氨肽酶、半纤维素酶及β-葡聚糖酶。
进一步的,所述复合酶中各单酶组分按重量份数计如下:中温α-淀粉酶5~8份,葡糖淀粉酶1~2份,普鲁兰酶0.5~1份,中性蛋白酶0.1~0.5份,氨肽酶1~5份,半纤维素酶5~10份,β-葡聚糖酶10~15份。
进一步的,所述辅料包括重量份数不大于77.4份的麦芽糊精。
具体的,上述用于提高和稳定啤酒发酵度的复合酶制剂包括按重量份数计的如下组分:中温α-淀粉酶5份,葡糖淀粉酶1份,普鲁兰酶0.5份,中性蛋白酶0.1份,氨肽酶1份,半纤维素酶5份,β-葡聚糖酶10份,及麦芽糊精77.4份。
具体的,上述用于提高和稳定啤酒发酵度的复合酶制剂包括按重量份数计的如下组分:中温α-淀粉酶6份,葡糖淀粉酶1.5份,普鲁兰酶0.8份,中性蛋白酶0.3份,氨肽酶3份,半纤维素酶8份,β-葡聚糖酶12份,及麦芽糊精68.4份。
具体的,上述用于提高和稳定啤酒发酵度的复合酶制剂包括按重量份数计的如下组分:中温α-淀粉酶8份,葡糖淀粉酶2份,普鲁兰酶1份,中性蛋白酶0.5份,氨肽酶5份,半纤维素酶10份,β-葡聚糖酶15份,及麦芽糊精58.5份。
进一步的,每克所述复合酶制剂中各单酶的酶活如下:中温α-淀粉酶10000U,葡糖淀粉酶酶活10万U,普鲁兰酶酶活3000U,中性蛋白酶酶活50万U,氨肽酶酶活1万U,β-葡聚糖酶酶活10万U,半纤维素酶酶活15万U。
另外,本发明还提供了一种上述复合酶制剂的应用,在啤酒糖化阶段,将上述复合酶制剂添加到由啤酒原料制成的待糖化的啤酒液中,所述复合酶制剂的添加量为啤酒原料总干重的0.01~0.1%。
进一步的,所述啤酒原料为麦芽,其制成待糖化的啤酒液的方法如下:取50g粉碎好的麦芽加入糖化杯中,加入400g的45℃温水,保温40分钟,然后以1℃/min的升温速率升至65℃,在65℃保温30分钟,然后同样速率升温到75℃,再保温20分钟后降到常温,过滤、灭菌制得麦汁。
进一步的,所述啤酒原料还包括配料,所述配料为大米、大麦、玉米淀粉中至少一种,所述配料的含量不超过麦芽重量的70%。
本发明中,复合酶制剂的各组分设计原则如下:
中温α-淀粉酶主要由优良菌株枯草芽孢杆菌(Bacillus subtilis)经过深层发酵、精制而成的高效酶制剂,最适温度是70~80℃,与糖化工艺条件吻合,可高效水解大麦中的糊化淀粉或破损淀粉中的α-1,4糖苷键,生成短链糊精和葡萄糖,配合葡糖淀粉酶、普鲁兰酶和内源酶β-淀粉酶的作用下,生成可发酵性糖,包含葡萄糖、果糖、麦芽糖、麦芽三糖。
葡糖淀粉酶是一种外切酶,由优良菌株黑曲霉(Aspergillus niger)经过深层发酵、精制而成的高效酶制剂,最适温度58~62℃,与糖化工艺条件吻合,可配合其他淀粉酶水解糊精非还原性末端生成葡萄糖,直接增加酵母可发酵性糖。
普鲁兰酶是一种脱支酶,由筛选的优良菌株地衣芽孢杆菌(Bacilluslicheniformis)经过深层发酵精炼而成,能够专一性催化水解支链淀粉支点α-1,6糖苷键,形成直链结构,最适温度55~65℃。由于酿造大麦啤酒时大麦中缺失该类酶,因此通过淀粉酶、葡糖淀粉酶和普鲁兰酶能够全面水解大麦中糊化的淀粉和破损淀粉,提高可发酵性糖的总量和其中葡萄糖的比例。
中性蛋白酶是一种由枯草芽孢杆菌(Bacillus subtilis)深层发酵精炼而成的蛋白酶,对蛋白底物具有广谱性,最适温度45~55℃,能够有效水解大麦蛋白中的肽键,降低蛋白分子量;氨肽酶是一种由优良菌株米曲霉发酵,经过生物现代技术精炼制成的酶制剂,是一种外切酶,最适温度50~55℃,能从多肽的N端逐个地切下氨基生成氨基酸,配合中性蛋白酶,能够有效生成小分子的氨基酸直接提供氮源,另一方面具有减少内切蛋白水解产生的苦味。
半纤维素酶是一组复杂酶的属名,一种戊聚糖,包含内切木聚糖酶、外切木聚糖酶、纤维二糖水解酶、阿拉伯呋喃糖苷酶等,是由优良菌株米曲霉发酵而成,最适温度48~52℃,可能够水解不溶性的阿拉伯木聚糖,降低溶液粘度,同时不溶性木聚糖的水解能够让淀粉和蛋白质释放,与相应的酶制剂结合更充分,增加水解效果。
β-葡聚糖酶是由优选菌株木霉深层发酵、精炼而得,包含β-1,3-D-葡聚糖水解酶,β-1,4-D-葡聚糖水解酶和β-1,3-1,4-D-葡聚糖水解酶,最适温度58~62℃,可以降低粘度,增加复合酶的水解效果,并生成少量的葡萄糖,从而增加啤酒发酵度。
啤酒作为以麦芽和水为主要原料、啤酒花为香料,经制麦芽、糖化、发酵等工序制成的富含营养物质和二氧化碳的酿造酒,具有独特的酒花香以及爽口苦味,深受消费者喜爱;糖化又称麦芽汁制备,通常指麦芽及辅料的粉碎,醪的糖化、过滤,以及麦汁煮沸、冷却的过程。由于本发明复合酶制剂主要用于改善啤酒糖化效果,促进酵母发酵,提高啤酒发酵度,发酵度则跟糖化后的麦汁直接相关,故本发明中,优选在啤酒糖化阶段将上述复配酶制剂添加于待糖化的啤酒液中;而为了更高的改善麦汁的糖化效果,可在啤酒糖化初期就将复配酶制剂添加于待糖化的麦汁中。
与现有技术相比,本发明的有益效果:
本发明提供的这种用于提高和稳定啤酒发酵度的复合酶制剂通过将不同淀粉酶和蛋白酶与木聚糖酶、葡聚糖酶和半纤维素酶进行复配后,能够有效稳定麦汁中的可发酵性糖和α-N的含量,其中淀粉酶有效降解麦芽中的淀粉成为可发酵性糖,蛋白酶则能随机水解大分子的蛋白质中的肽键,配合外切蛋白酶生成α-氨基酸、二肽或三肽等短链蛋白质从而提供氮源,而配合木聚糖酶、葡聚糖酶和半纤维素酶能够有效地增加淀粉酶和蛋白酶的水解效果,同时也能将一些戊聚糖、葡聚糖、半纤维素等水解成小分子的可发酵糖,通过酶制剂各单酶之间的协同效果,有效地降解原料中破损淀粉颗粒、糊化的淀粉、蛋白质和糖蛋白,以及少量葡聚糖糖、半纤维素、木聚糖等,效果明显优于单酶效果,同时选择麦芽糊精辅料使得酶制剂的混合更加均匀且提高了可发酵性糖的含量;另外,该复合酶制剂准备方法简单,由于都是粉剂,复合酶制剂各组分相互之间不作用,制备简单且稳定,将该酶应用于糖化阶段后,有效解决麦芽不稳定的因素,解决发酵度低或差异大的问题。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其它实施例,都属于本发明保护的范围。
下述实施例未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
实施例1:
本实施例提供了一种用于提高和稳定啤酒发酵度的复合酶制剂,由按重量份数计的如下组分混合均匀制得:中温α-淀粉酶5份,葡糖淀粉酶1份,普鲁兰酶0.5份,中性蛋白酶0.1份,氨肽酶1份,半纤维素酶5份,β-葡聚糖酶10份,及麦芽糊精77.4份。
其中,每克所述复合酶制剂中各单酶的酶活如下:中温α-淀粉酶10000U,葡糖淀粉酶酶活10万U,普鲁兰酶酶活3000U,中性蛋白酶酶活50万U,氨肽酶酶活1万U,β-葡聚糖酶酶活10万U,半纤维素酶酶活15万U。
实施例2:
本实施例提供了一种用于提高和稳定啤酒发酵度的复合酶制剂,由按重量份数计的如下组分混合均匀制得:中温α-淀粉酶6份,葡糖淀粉酶1.5份,普鲁兰酶0.8份,中性蛋白酶0.3份,氨肽酶3份,半纤维素酶8份,β-葡聚糖酶12份,及麦芽糊精68.4份。
其中,每克所述复合酶制剂中各单酶的酶活如下:中温α-淀粉酶10000U,葡糖淀粉酶酶活10万U,普鲁兰酶酶活3000U,中性蛋白酶酶活50万U,氨肽酶酶活1万U,β-葡聚糖酶酶活10万U,半纤维素酶酶活15万U。
实施例3:
本实施例提供了一种用于提高和稳定啤酒发酵度的复合酶制剂,由按重量份数计的如下组分混合均匀制得:中温α-淀粉酶8份,葡糖淀粉酶2份,普鲁兰酶1份,中性蛋白酶0.5份,氨肽酶5份,半纤维素酶10份,β-葡聚糖酶15份,及麦芽糊精58.5份。
其中,每克所述复合酶制剂中各单酶的酶活如下:中温α-淀粉酶10000U,葡糖淀粉酶酶活10万U,普鲁兰酶酶活3000U,中性蛋白酶酶活50万U,氨肽酶酶活1万U,β-葡聚糖酶酶活10万U,半纤维素酶酶活15万U。
取5个不同批次的麦芽作为啤酒原料,将上述实施例1~3分别按照麦芽干重的0.01%添加于待糖化的麦汁中,经过糖化,定容、灭菌,接种,测定最终发酵度,并设定空白对照组,测定结果如表1所示,增长率为相对空白对照的增长率。其中,麦芽糖化处理方式为:取50g粉碎好的麦芽加入糖化杯中,加入400g的45℃温水,保温40分钟,然后以1℃/min的升温速率升至65℃,在65℃保温30分钟,然后同样速率升温到75℃,然后保温20分钟后降到常温,过滤、灭菌制的麦汁。啤酒发酵度或麦芽极限发酵度的检测方法为:取200mL上述麦汁,加入10g抽干后的酵母饼,震荡几次,用纱布封好,于25℃全温震荡发酵24h,加入2g硅藻土到过滤装置对发酵液过滤,对滤液震荡除气,将过滤除气后的发酵液放入Anton Paar啤酒分析仪中检测其发酵度。
表1:不同批次麦芽的发酵度
由表1可知,添加本发明的复合酶制剂,对于发酵度低的麦芽改善效果更好,因此,使用本发明的复合酶制剂可以降低啤酒发酵度的偏差波动,提高麦芽批次间的稳定性;此外,添加此复合酶制剂可以提高啤酒发酵度。
实施例4:
本实施例取同一批次的麦芽作为啤酒原料,选用不同淀粉酶作实验组按照相同性价比以麦芽干重的0.02%添加于待糖化的麦汁中,经过糖化,定容、灭菌,接种,测定最终发酵度,并设定空白对照组,麦芽糖化处理方式及啤酒发酵度的测定方法同上述实施例1,测定结果如表2所示。其中,选择中温α-淀粉酶,高温α-淀粉酶,真菌α-淀粉酶,葡糖淀粉酶,普鲁兰酶和β-淀粉酶分别为实验组1~6,增长率为相对空白对照的增长率。
表2:不同淀粉酶对啤酒发酵度的影响
由表2可知,本发明复合酶制剂中所选用的淀粉酶通过筛选大量不同淀粉酶而得到的,而其中中温α-淀粉酶、葡糖淀粉酶和普鲁兰酶对提高啤酒发酵度具有更好效果,因此,本发明复合酶制剂中淀粉酶选择中温α-淀粉酶、葡糖淀粉酶和普鲁兰酶。
实施例5:
本实施例取同一批次的麦芽作为啤酒原料,选用不同蛋白酶作实验组按照相同性价比以麦芽干重的0.005%添加于待糖化的麦汁中,经过糖化,定容、灭菌,接种,测定最终发酵度,并设定空白对照组,麦芽糖化处理方式及啤酒发酵度的测定方法同上述实施例1,测定结果如表3所示。其中,选择酸性蛋白酶、碱性蛋白酶、中性蛋白酶、木瓜蛋白酶和氨肽酶分别为实验组1~5,增长率为相对空白对照的增长率。
表3:不同蛋白酶对啤酒发酵度的影响
由表3可知,本发明复合酶制剂中所选用的蛋白酶通过筛选大量不同蛋白酶而得到的,而其中中性蛋白酶和氨肽酶对提高啤酒发酵度具有更好效果,因此,本发明复合酶制剂中蛋白酶选择中性蛋白酶和氨肽酶。
实施例6:
本实施例取同一批次的麦芽作为啤酒原料,选用不同非淀粉多糖酶作实验组按照相同性价比以麦芽干重的0.02%添加于待糖化的麦汁中,经过糖化,定容、灭菌,接种,测定最终发酵度,并设定空白对照组,麦芽糖化处理方式及啤酒发酵度的测定方法同上述实施例1,测定结果如表4所示。其中,选择半纤维素酶、β-葡聚糖酶、甘露聚糖酶、半乳糖苷酶和纤维素酶分别为实验组1~5,增长率为相对空白对照的增长率。
表4:不同非淀粉多糖酶对啤酒发酵度的影响
由表4可知,本发明复合酶制剂中所选用的非淀粉多糖酶通过筛选大量不同非淀粉多糖酶而得到的,而其中半纤维素酶和β-葡聚糖酶对提高啤酒发酵度具有更好效果,因此,本发明复合酶制剂中非淀粉多糖酶选择半纤维素酶和β-葡聚糖酶。
以上例举仅仅是对本发明的举例说明,并不构成对本发明的保护范围的限制,凡是与本发明相同或相似的设计均属于本发明的保护范围之内。
Claims (10)
1.一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,包括复合酶及辅料,所述复合酶包括如下单酶组分:中温α-淀粉酶、葡糖淀粉酶、普鲁兰酶、中性蛋白酶、氨肽酶、半纤维素酶及β-葡聚糖酶。
2.如权利要求1所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,所述复合酶中各单酶组分按重量份数计如下:中温α-淀粉酶5~8份,葡糖淀粉酶1~2份,普鲁兰酶0.5~1份,中性蛋白酶0.1~0.5份,氨肽酶1~5份,半纤维素酶5~10份,β-葡聚糖酶10~15份。
3.如权利要求2所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,所述辅料包括重量份数不大于77.4份的麦芽糊精。
4.如权利要求3所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,包括按重量份数计的如下组分:中温α-淀粉酶5份,葡糖淀粉酶1份,普鲁兰酶0.5份,中性蛋白酶0.1份,氨肽酶1份,半纤维素酶5份,β-葡聚糖酶10份,及麦芽糊精77.4份。
5.如权利要求3所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,包括按重量份数计的如下组分:中温α-淀粉酶6份,葡糖淀粉酶1.5份,普鲁兰酶0.8份,中性蛋白酶0.3份,氨肽酶3份,半纤维素酶8份,β-葡聚糖酶12份,及麦芽糊精68.4份。
6.如权利要求3所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,包括按重量份数计的如下组分:中温α-淀粉酶8份,葡糖淀粉酶2份,普鲁兰酶1份,中性蛋白酶0.5份,氨肽酶5份,半纤维素酶10份,β-葡聚糖酶15份,及麦芽糊精58.5份。
7.如权利要求1所述的一种用于提高和稳定啤酒发酵度的复合酶制剂,其特征在于,每克所述复合酶制剂中各单酶的酶活如下:中温α-淀粉酶10000U,葡糖淀粉酶酶活10万U,普鲁兰酶酶活3000U,中性蛋白酶酶活50万U,氨肽酶酶活1万U,β-葡聚糖酶酶活10万U,半纤维素酶酶活15万U。
8.如权利要求1~7任一项所述的用于提高和稳定啤酒发酵度的复合酶制剂的应用,其特征在于,在啤酒糖化阶段,将权利要求1~7任一项所述的复合酶制剂添加到由啤酒原料制成的待糖化的啤酒液中,所述复合酶制剂的添加量为啤酒原料总干重的0.01~0.1%。
9.如权利要求8所述的复合酶制剂的应用,其特征在于,所述啤酒原料为麦芽,其制成待糖化的啤酒液的方法如下:取50g粉碎好的麦芽加入糖化杯中,加入400g的45℃温水,保温40分钟,然后以1℃/min的升温速率升至65℃,在65℃保温30分钟,然后同样速率升温到75℃,再保温20分钟后降到常温,过滤、灭菌制得麦汁。
10.如权利要求9所述的复合酶制剂的应用,其特征在于,所述啤酒原料还包括配料,所述配料为大米、大麦、玉米淀粉中至少一种,所述配料的含量不超过麦芽重量的70%。
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