CN115413779B - 一种通过微生物发酵加速陈皮陈化的方法 - Google Patents
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Abstract
本发明公开了卷枝毛霉(Mucor circinelloides)在加速陈皮陈化中的应用,本发明还公开了一种通过微生物发酵加速陈皮陈化的方法,利用卷枝毛霉对陈皮黄酮类物质的高效转化能力促进陈皮的陈化,通过发酵能够在极短的时间内将橘皮转化为陈皮,从而丰富了陈皮微生物陈化的菌种资源,提高了陈化效率,具有良好的应用前景。
Description
技术领域
本发明涉及陈皮的陈化工艺,具体涉及一种通过微生物发酵对陈皮进行陈化的方法。
背景技术
陈皮为芸香科植物橘(Citrus Reticulate Blabco)干燥成熟果皮,能够调节中气、化痰祛湿热等,是我国传统的药食两用药材。药材中陈皮分为“陈皮”和“广陈皮”,其中道地的广陈皮来源于广东省新会地区的茶枝柑,质地良好,具有独特的药用价值,在广东盛传“百年陈皮千年参,一两陈皮一两金”,素有“陈久者良”的说法。现代药理学研究表明陈皮含有丰富的黄酮类化合物,具有良好的抗炎、抗氧化、抗癌及抗病毒等作用。
陈皮需陈化三年及以上才能入药。自2000年以来,新会区着重新会陈皮品牌打造,使陈皮产业迎来快速发展,影响力从地方走向全国乃至国际市场,产量供不应求。在陈皮陈化过程中,药效物质黄酮类化合物,如橙皮苷、川陈皮素、橘皮素、多甲氧基黄酮等含量逐渐增加,挥发油物质慢慢转化,最终形成陈皮独特的怡人香气和功效。现今陈皮陈化工序以自然陈化为主,即储存在通风良好的陈化仓中自然陈化三到五年,陈化时间过长,不利于提高陈皮产量,且自然储藏环境复杂不可控制,难以实现标准化,而且还易造成陈皮的霉变腐烂或虫蛀。为了缓解陈皮供求紧张的现状、标准化陈皮品质和简化陈化工序具有重要意义。
目前加速陈皮陈化的方法主要有物理方法和微生物方法,物理方法为调控陈化环境温度、湿度和光照;微生物方法为引入微生物发酵陈化。CN105581288A公开了一种陈皮的快速陈化方法,具体是将陈皮在高温条件下与自然条件下交替陈化,如此反复,能使陈化总时间缩短到20-90天;CN111067956A公开了一种加速陈皮陈化的方法,具体是将晒干的柑皮置于湿度90%-95%的条件下,保持2-5天,取出后将柑皮置于湿度70%-75%的条件下,保持1-4天,如此交替反复。以上两种方法虽能对陈皮陈化起到一定程度上的加速效果,但对陈化环境要求较高,加工时间相对较长,步骤繁琐,不方便推广。CN111973673A的专利公开了一种加快陈皮陈化的方法,具体是喷洒益生菌细菌发酵液至柑皮上,发酵14-28天后干燥。但益生菌资源日趋紧张,因此有必要开发更多的菌种资源。真菌种类繁多且在自然界广泛存在,近年来,真菌在陈皮陈化中所发挥的作用逐渐被发现,但目前尚没有利用卷枝毛霉(Mucor circinelloides)加速陈皮陈化的报导。
发明内容
本发明的目的在于克服上述现有技术的不足,提供一种通过微生物发酵对陈皮进行陈化的方法,旨在为陈皮的微生物陈化提供更多的菌种资源,同时进一步缩短陈化周期。
上述目的是通过以下技术方案实现的:
一种通过微生物发酵加速陈皮陈化的方法,包括以下步骤:
(1)将茶枝柑洗净剥皮,将果皮干燥后粉碎成细粉,然后紫外灭菌;
(2)将卷枝毛霉在培养基中进行培养,然后取孢子加至无菌水中,配制成孢子悬浮液;
(3)向灭菌后的果皮粉中喷洒孢子悬浮液,喷洒量为果皮粉重量的0.1-3%,搅拌混匀,调节果皮粉含水量为50-90%,然后进行渥堆发酵,发酵温度为45-55℃,时间7-14天;
(4)发酵结束后将果皮粉低温干燥。
优选地,所述培养基是PDA固体培养基。
优选地,所述孢子悬浮液的浓度是105-109CFU/mL。
优选地,所述孢子悬浮液的喷洒量为果皮粉重量的0.1-1%。
优选地,所述含水量为70-80%,在含水量范围内,真菌代谢活动强,对陈皮中黄酮类物质转化效率高。
优选地,所述发酵温度为50℃。
本发明还提供了卷枝毛霉在加速陈皮陈化中的应用。
本发明的有益效果是:
本发明利用卷枝毛霉对陈皮黄酮类物质的高效转化能力促进柑皮的陈化,能够在极短的时间内将橘皮转化为陈皮,从而丰富了陈皮微生物陈化的菌种资源,提高了陈化效率,具有良好的应用前景。
具体实施方式
下面结合具体实施例对本发明进行详细说明。以下各实施例中所使用的原料茶枝柑均为同一公司同一批次的产品,所使用的菌种如卷枝毛霉菌、黑曲霉菌、红酵母菌、小孢根霉菌均为市售产品。
实施例1
(1)选取成色好,无明显病虫害的茶枝柑为原料,洗净剥皮,将果皮干燥后粉碎成细粉,然后紫外照射灭菌30min;
(2)称取马铃薯葡萄糖琼脂(PDA)培养基46g,溶解于1000mL水中,在121℃条件下灭菌15min,倒平板凝固,得真菌固体培养基;将卷枝毛霉接种到上述固体培养基,28℃培养箱培养3天,然后用接种环取孢子至无菌水中,配制成浓度为106CFU/mL的孢子悬浮液;
(3)向灭菌后的果皮粉中喷洒孢子悬浮液,喷洒量为果皮粉重量的0.5%,边喷洒边搅拌混匀,接着喷洒无菌水控制果皮粉含水量为75%,然后进行渥堆发酵,发酵温度为50℃,时间10天,发酵过程中定期翻动并通风;
(4)发酵结束后将果皮粉在50-60℃下干燥,至含水量低于10%即可,将干燥后的产物用玉米纤维茶袋包装,置于阴凉干燥处储存。
实施例2
(1)将茶枝柑洗净剥皮,将果皮干燥后粉碎成细粉,然后紫外灭菌;
(2)将卷枝毛霉接种到PDA固体培养基进行培养,然后取孢子至无菌水中,配制成浓度为106CFU/mL的孢子悬浮液;
(3)向灭菌后的果皮粉中喷洒孢子悬浮液,喷洒量为果皮粉重量的0.1%,边喷洒边搅拌混匀,接着喷洒无菌水控制果皮粉含水量为80%,然后进行渥堆发酵,发酵温度为55℃,时间14天,发酵过程中定期翻动并通风;
(4)发酵结束后将果皮粉低温干燥至含水量低于10%,将干燥后的产物包装后置于阴凉干燥处储存。
实施例3
(1)将茶枝柑洗净剥皮,将果皮干燥后粉碎成细粉,然后紫外灭菌;
(2)将卷枝毛霉接种到PDA固体培养基进行培养,然后取孢子至无菌水中,配制成浓度为106CFU/mL的孢子悬浮液;
(3)向灭菌后的果皮粉中喷洒孢子悬浮液,喷洒量为果皮粉重量的1%,边喷洒边搅拌混匀,接着喷洒无菌水控制果皮粉含水量为60%,然后进行渥堆发酵,发酵温度为45℃,时间7天,发酵过程中定期翻动并通风;
(4)发酵结束后将果皮粉低温干燥至含水量低于10%,将干燥后的产物包装后置于阴凉干燥处储存。
试验例1感官评价试验
邀请20名感官评价员对以上实施例制得的陈皮粉进行感官评价,评价时取1g陈皮粉加50ml开水泡茶,茶汤评分表如表1所示,感官评价结果如表2所示。
表1感官评分表
表2感官评价结果
样品 | 感官评价得分 |
实施例1 | 88 |
实施例2 | 86 |
实施例3 | 81 |
对照组1 | 55 |
对照组2 | 84 |
对照组1是未经陈化的新鲜橘皮粉,对照组2是自然陈化三年的陈皮经粉碎后得到的陈皮粉。
结果表明,陈皮粉冲泡口感佳,香味醇厚,口感优于橘皮粉。使用卷枝毛霉发酵陈化的陈皮粉其感官评分与自然陈化相当。
试验例2橙皮苷、川陈皮素、橘皮素含量测定
在橘皮陈化过程中,主要成分橙皮苷含量逐渐下降,川陈皮素、橘皮素含量不断上升,颜色逐渐变暗,质地变薄。通过对实施例中的橙皮苷、川陈皮素、橘皮素含量进行检测比较,能更加直观地证明陈化效果。
试验过程:橙皮苷、川陈皮素和橘皮素含量通过超高效液相色谱方法测定。具体细节如下:采用超高效液相色谱(UPLC)测定黄酮类化合物含量:色谱柱为Waters ACQUITYHSS C18色谱柱(2.1×100mm i.d.,1.8μm),流动相A相为乙腈,B相为0.1%甲酸的水溶液(梯度洗脱)。流速为0.2mL/min,柱温为25℃,样品进样量为5μL。检测波长为283nm(橙皮苷)和330nm(川陈皮素、橘皮素)。
试验结果如表3所示。
表3实施例与对比例陈皮粉有效物质含量变化(%)
样品 | 橙皮苷 | 川陈皮素 | 橘皮素 |
实施例1 | 4.32 | 0.27 | 0.17 |
实施例2 | 4.36 | 0.26 | 0.15 |
实施例3 | 4.34 | 0.25 | 0.17 |
对比例1 | 4.54 | 0.19 | 0.08 |
对比例2 | 4.48 | 0.22 | 0.10 |
对比例3 | 4.42 | 0.20 | 0.13 |
对照组 | 4.21 | 0.29 | 0.16 |
其中,对比例1、2、3是分别用黑曲霉(Aspergillus niger)、红酵母(Rhodotorula)、小孢根霉(Rhizopus microsporus)代替卷枝毛霉进行渥堆发酵制备的陈皮粉,具体的工艺参数和方法与实施例1相同;对照组是自然陈化三年的陈皮粉。
陈化年限越久,陈皮药用功效越显著,香味越浓,价格越高。陈化过程主要是由于微生物代谢活动一方面使其药效成分含量增加,另一方面使其质地变薄,药效物质更易析出。分析上表可以看出,本发明提供的加速工艺中橙皮苷含量较低,川陈皮素和橘皮素含量较高,效果与对照组接近。黑曲霉、红酵母、小孢根霉三种真菌也具有加速陈皮陈化的作用,但效果不如卷枝毛霉。
综上所述,本发明提供了一种陈皮粉加速陈化方法,该方法选用卷枝毛霉为接种菌,通过渥堆发酵,大幅缩短了陈皮陈化时间,达到了与自然陈化相近的效果。
最后应当说明的是,以上实施例仅用以解释本发明的技术方案而非对本发明保护范围的限制,本领域技术人员在不违背本发明创造精神的前提下还可作出各种等同的变型或替换,这些等同的变型或替换均包含在本申请权利要求所限定的范围内。
Claims (7)
1.卷枝毛霉(Mucor circinelloides)在加速陈皮陈化中的应用。
2.一种通过微生物发酵加速陈皮陈化的方法,其特征在于包括以下步骤:
(1)将茶枝柑洗净剥皮,将果皮干燥后粉碎成细粉,然后紫外灭菌;
(2)将卷枝毛霉在培养基中进行培养,然后取孢子加至无菌水中,配制成孢子悬浮液;
(3)向灭菌后的果皮粉中喷洒孢子悬浮液,喷洒量为果皮粉重量的0.1-3%,搅拌混匀,调节果皮粉含水量为50-90%,然后进行渥堆发酵,发酵温度为45-55℃,时间7-14天;
(4)发酵结束后将果皮粉低温干燥。
3.如权利要求2所述通过微生物发酵加速陈皮陈化的方法,其特征在于:所述培养基是PDA固体培养基。
4.如权利要求2所述通过微生物发酵加速陈皮陈化的方法,其特征在于:所述孢子悬浮液的浓度是105-109CFU/mL。
5.如权利要求2所述通过微生物发酵加速陈皮陈化的方法,其特征在于:所述孢子悬浮液的喷洒量为果皮粉重量的0.1-1%。
6.如权利要求2所述通过微生物发酵加速陈皮陈化的方法,其特征在于:所述含水量为70-80%。
7.如权利要求2所述通过微生物发酵加速陈皮陈化的方法,其特征在于:所述发酵温度为50℃。
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