CN1061094C - 淡水珍珠粉末的生物降解方法 - Google Patents
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Abstract
本发明涉及一种生物工程技术,用微生物发酵方法降解淡水珍珠粉末的方法。该方法所用的菌种为科植1号菌,一次发酵把珍珠粉末的碳酸钙和蛋白完全水解,是一种效率高,成本低,适用于工业化生产的水解珍珠粉的技术,制得口感好,质量高的珍珠生物降解液,适合作为医药、保健品、食品、日用化工的原料。
Description
本发明涉及一种生物学技术,用微生物发酵方法降解水珍珠粉末。
珍珠是重要的药材,在我国已有两千年的应用的历史。历代医药古籍有珍珠“明目、翳肤、止泄”作用的记载。它对内科、儿科、眼科、外科和妇产科某些疾病具有特殊功效,这已被临床医学所确认。以珍珠为主料的方剂已被收入《中华人民共和国药典》。
近代科学研究查明珍珠由两类化合物构成,一类是角壳硬蛋白约占珍珠重量的4%,另一类是蚌类从生长水体中富集的无机元素,主要是钙38.82%,碳12.57%,氢0.34%,氮0.52%。同时还含有各种微量元素和痕量元素如:镁、锰、铜、铝、铁、钨、锌、钡、铬、镍、钴、银、金、钛、硅、锂、碘、溴、钍、镧、钪、硒等,此外还有约1%的水分。角壳硬蛋白和人体必需的各种微量和超微量元素可能是珍珠疗效的主要成份。
珍珠粉末是不溶于水的粉笔灰状的不定型固体,有效成份难溶于水,很久以来一直只能作为中药的材料,近年来由于珍珠产量的增加和人民生活水平的提高,国内外都研究拓展珍珠利用的新领域,除了传统的药用之外,在保健饮料(KICCHO SHuZO KK(KICC-)Drugeffective processedalcoholic drinks-Contg.peasl from eitherfresh or sea waterJP05192126)、口服液或胶囊(陈国方等,珍珠口服液及其生产工艺,中国专利CN1091008A),日用化工产品生产方面(Pola CHEM IND INC(POKK),Bathing agentpromoting blood Circulation-Cotains pearlcnlture extract obtd.by hydrolysing pesrl Oystes shell or pesrls,JP05148131)也进行了很有特色的研究(陈国方等,珍珠口服液及其生产工艺,中国专利CN1091008A)最关键的技术就是把水不溶性的珍珠粉末液化。这方面研究工作主要有两个类型:一是用无机酸(盐酸、磷酸或硫酸等)或有机酸(醋酸或乳酸)将珍珠的碳酸钙溶解,再将蛋白质加盐酸回流水解,转化成氨基酸(潘炳炎等,珍珠实用新技术,中国农业科技出版社1994年;卢济台等,用珍珠或珍珠层粉制取全珍珠方法,中国专利CN1107044A);另一种方法称为“双解法”,即用酸溶解碳酸钙,然后用木瓜蛋白酶水解珍珠蛋白质(李新建等,由固体珍珠制备液体珍珠的方法,中国专利CN1063896A;黄永昌等,酶介珍珠液,中国专利CN1107335A)。第一类方法不足之处在于:加入强酸后产生大量二氧化碳以气泡的形式溢出,使反应难以控制,此外这个过程还损失了部分有效成份,酸水解后所得溶液实际上是氨基酸,这些氨基酸是否是珍珠的有效成份尚待证实。第二种方法除了用酸去钙与前一种方法有同样不足之外,增加了过滤、洗涤,加酶分解,再过滤等多种步骤,给工业化生产带来不便,不过用酶水解比酸解温和得多,可能保留较多的有效成份。但这两种方法的产物中存在氯化钙等成分,口感不好,有苦涩味。不符合饮料生产的要求。
本工作的目的是提供一种生物工程技术,利用本研究组分离得到的菌株,一次发酵把珍珠的碳酸钙和角壳蛋白水解,得到效率高、成本低、适于工业化生产的珍珠水解技术出制造口感好,高质量的珍珠生物降解液,适合作为医药、食品、日用化工的原料。
生物降解珍珠粉末的工艺流程和步骤如下:
1)培养科植1号酵母菌体
2)水解珍珠粉末
3)分离水解液和菌体得到纯珍珠粉水解液
降解珍珠粉末所用菌种为酿酒酵母(Saccharomyces cerevisiae)
自行分离得到的并命名为科植1号酵母菌,已于1996年9月5日在中国微生物菌种保藏管理委员会普通微生物中心(CGMCC,地址:中国,北京,中关村,邮政编码:100080)进行了保藏,编号为CGMCC NO.0273。
科植1号酵母菌种先通过斜面培养基,接着进行摇瓶培养,然后种子罐培养,得到发酵菌种。
降解珍珠粉末的方法是通过将培养好的科植1号酵母菌接入大罐进行培养发酵,同时加入珍珠粉末,在一定的条件下进行水解。菌种培养和水解的具体条件如下所述:
培养菌种一般用固体培养基斜面养也可用培养皿培养,土豆培养基,蔗糖2%,磷酸二氢钾0.15-0.5%,硫酸镁0.05%-0.15%,马铃薯浸出物15-20%,摇瓶培养种子罐发酵与斜面培养基相同。大罐发酵培养基用蔗糖1-2%,土豆浸出液15-20%,同时放入100-200目珍珠粉0.25-0.8%。培养发酵一般温度25℃-35℃,16-24小时。PH6.0-7.5。
最佳实施例:
步骤1:斜面培养菌种
扩大生产需要的原种,科植1号菌种的培养基配方为:(克/升)
蔗糖 20克
磷酸二氢钾 1.5克
硫酸镁 0.5克
马铃薯浸出物 200克
洋菜粉末 20克
取新鲜马铃薯洗净去皮称取200克加水200毫升煮沸30分钟,过滤取清液备用。
上述成份充分溶解后分别装入直径13×150mm试管中,加封口膜后在高压锅内灭菌120℃20分钟,压力平衡后取出,一端置于高处,凝固后即为斜面。制备好的斜面包好放在冰箱内备用,使用量在无菌室划线接种,在25℃-35℃温箱内培养12-24小时即可使用。
步骤2:摇瓶培养(扩大培养)
250-300毫升三角瓶,每瓶加入100毫升培养基,(克/升)
蔗糖 20克
磷酸二氢钾 1.5克
硫酸镁 0.5克
马铃薯浸出物 150毫升
定容至1000毫升,分装成每瓶100毫升,加封口膜,高压灭菌120℃20分钟,平衡后取出在无菌室接种,摇床振培养35℃,频率150次/分,培养24时。
步骤3种子罐发酵
种子罐发酵培养,灭菌冷却至35℃,接种摇瓶发酵液1.5升(即5%)。并通入无菌空气,相对通风量为1∶0.5,即30升种子缸每分钟通无菌空气15升。保持发酵温度25℃-35℃,24小时完成扩大培养接种至大罐。
步骤4大罐发酵
发酵培养基为土豆浸出液15%,蔗糖2%,加入0.6-0.8%珍珠粉,高温消毒培养基及珍珠粉,冷却至35℃后接入种子罐发酵液,相对通风量为1∶0.5,保持温度35℃并搅拌,12-16小时停止发酵,加蒸气至100℃保温半小时以杀灭菌体。
步骤5过滤
发酵液经硅藻土滤机过滤,除去固体组份,得到浅琥珀色的清液即为珍珠水解液。
和现有的珍珠水解技术相比本方法有三个主要的优点:
1、在珍珠水解阶段,培养基中只有两种天然产品,不加入任何人工合成的化学物质,无污染物或其他有害成份。
2、工艺简便有效,每生产500斤珍珠水解液只需人民币300元,珍珠水解率达80%。
3、珍珠水解在PH6-7,温度37℃下进行,保持了较多的珍珠有效成份,水解液含钙1.16克/升,蛋白质1-2克/升,蛋白质水解产物含17种氨基酸,人类必需的八种氨基酸含量丰富。水解液中的钙部分转变为有机钙或大分子钙,所以口感很好,是较好的制造饮料的原料。
本生产工艺适用于珍珠深加工和综合利用,以珍珠生物降解为关键技术,生产系列珍珠产品,如超细可溶珍珠粉,水解液经浓缩并与中药配伍可制成药用珍珠胶囊,与果汁或其他材料配合可制成珍珠饮料,与油脂配合可制成各种化妆品,皮肤清洁品。
Claims (4)
1.一种淡水珍珠粉末的降解方法,包括如下步骤:
1)培养酿酒酵母科植1号酵母菌株CGMCC 0273;
2)将珍珠粉末加入上述酵母的培养物中进行水解;
3)分离水解液和菌体得到纯珍珠粉水解液。
2.按照权利要求1所述的方法,其特征在于,用于培养科植1号酵母菌株CGMCC 0273的培养基包含下述成份:
蔗糖 1-2%;
磷酸二氢钾 0.15-0.5%;
硫酸镁 0.05-0.15%;
马铃薯浸出物 15-20%;
洋菜粉末 1-2%。
3.按照权利要求2所述的方法,其特征在于,所述的培养基包含下述成份:
蔗糖 10-20克/升
磷酸二氢钾 1.5克/升
硫酸镁 0.5克/升
马铃薯浸出物 200克/升
洋菜粉末 20克/升
4.按照权利要求1所述的方法,其特征在于,发酵水解珍珠粉的培养基包含下述成份:
蔗糖 1-2%
土豆浸出液 15-20%
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CN105769725A (zh) * | 2016-03-02 | 2016-07-20 | 禹州市天源生物科技有限公司 | 一种迷迭香保湿水及其制作方法 |
CN106265412B (zh) * | 2016-08-04 | 2019-03-26 | 海南京润珍珠生物技术股份有限公司 | 一种利用益生菌发酵制备珍珠水解液的方法 |
CN112569172A (zh) * | 2019-09-29 | 2021-03-30 | 欧诗漫生物股份有限公司 | 珍珠制备品及其制备方法和用途 |
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CN1051394A (zh) * | 1990-12-06 | 1991-05-15 | 王四明 | 水溶性全成分珍珠粉系列产品新制备方法 |
CN1063896A (zh) * | 1991-02-04 | 1992-08-26 | 李新建 | 由固体珍珠制备液体珍珠的方法 |
CN1080323A (zh) * | 1993-03-05 | 1994-01-05 | 何亚娟 | 珍珠原液及其制备方法(双解法) |
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CN1051394A (zh) * | 1990-12-06 | 1991-05-15 | 王四明 | 水溶性全成分珍珠粉系列产品新制备方法 |
CN1063896A (zh) * | 1991-02-04 | 1992-08-26 | 李新建 | 由固体珍珠制备液体珍珠的方法 |
CN1080323A (zh) * | 1993-03-05 | 1994-01-05 | 何亚娟 | 珍珠原液及其制备方法(双解法) |
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