CN113662181B - 具有抗疲劳功能活性的马尾藻水解物及其制备方法与应用 - Google Patents
具有抗疲劳功能活性的马尾藻水解物及其制备方法与应用 Download PDFInfo
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- CN113662181B CN113662181B CN202111238077.9A CN202111238077A CN113662181B CN 113662181 B CN113662181 B CN 113662181B CN 202111238077 A CN202111238077 A CN 202111238077A CN 113662181 B CN113662181 B CN 113662181B
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Abstract
本发明公开了一种具有抗疲劳功能活性的马尾藻水解物及其制备方法与应用，属于生物工程技术领域，制备该马尾藻水解物的方法为：将马尾藻粉碎，加水配制马尾藻悬浮液；加入褐藻胶裂解酶ALVC‑03，酶解；酶解后加入葡萄糖，高压灭菌，接种枯草芽孢杆菌，发酵；发酵结束后高压灭菌，离心，上清液即为具有抗疲劳功能活性的马尾藻水解物；所述褐藻胶裂解酶ALVC‑03的氨基酸序列如SEQ ID NO:1所示。本发明的马尾藻水解物具有抗疲劳功能活性，能够缓解过量运动导致的机体疲劳，使机体耐力和力量得到有效恢复，具有成为缓解过量运动疲劳的功能食品原料的应用潜力，可以以该马尾藻水解物为原料制备缓解运动疲劳的功能性食品或饮料。
Description
技术领域
本发明涉及一种具有抗疲劳功能活性的马尾藻水解物，及其制备方法与应用，属于生物工程技术领域。
背景技术
马尾藻分布广泛，是一种重要的经济藻类。但由于马尾藻有着坚固的细胞壁结构，导致其作为食品原料时具有消化利用率低、功能活性难以发挥等缺点，因此需要对马尾藻进行破壁和生物转化处理，使其细胞中的活性成分得以释放，同时经过生物转化，使其功能活性进一步提升。
随着现代社会人们对生活品质的追求，越来越多的人开始参与到运动与健身中，但很多人因为缺乏相应的专业知识，往往会过量运动从而引起疲劳，导致耐力和力量下降等，反而会导致身体状态和生活质量下降。开发一种能够抗运动疲劳的功能性食品原料，是解决这一问题的重要途径。
利用天然藻类为原料，通过生物转化方法释放藻中的有效成分，开发成为具有抗疲劳活性的功能食品原料，是海藻高值化利用的重要途径。
发明内容
针对上述现有技术，本发明提供了一种具有抗疲劳功能活性的马尾藻水解物，还提供了其制备方法，以及在制备功能性食品或饮料中的应用。本发明还提供了一种褐藻胶裂解酶及其编码基因。
本发明是通过以下技术方案实现的：
一种制备马尾藻水解物的方法：将马尾藻粉碎，加水配制马尾藻悬浮液；加入褐藻胶裂解酶ALVC-03或含褐藻胶裂解酶ALVC-03的酶液，酶解；酶解后加入葡萄糖，高压灭菌，接种枯草芽孢杆菌，发酵；发酵结束后高压灭菌，离心，上清液即为具有抗疲劳功能活性的马尾藻水解物；所述褐藻胶裂解酶ALVC-03，其氨基酸序列如SEQ ID NO:1所示。
进一步地，所述马尾藻悬浮液，马尾藻与水的质量体积比为1:3～8（单位g:ml），优选1:5。
进一步地，所述褐藻胶裂解酶ALVC-03的质量为马尾藻质量的3%～8%，优选5%。
进一步地，所述酶解的条件为：温度35～45℃，pH 6.5～7.5，时间8～12小时。优选的，温度40℃，pH 7.0，时间10小时。
进一步地，所述葡萄糖的添加量为酶解液体积的0.5%～2%（单位g/ml，即：每100ml酶解液中加入0.5%～2g的葡萄糖），优选1%。
进一步地，所述高压灭菌的条件为：121℃灭菌20 min。
进一步地，所述枯草芽孢杆菌，是保藏编号为CGMCC 1.15792的枯草芽孢杆菌。
进一步地，所述发酵的条件为：35～40℃发酵25～35小时。优选的，37℃发酵30小时。
进一步地，所述离心的条件为：8000 r/min离心10 min。
利用上述方法制备得到的马尾藻水解物，具有抗疲劳功能活性，可以以该马尾藻水解物为原料制备缓解运动疲劳的功能性食品或饮料。
一种褐藻胶裂解酶ALVC-03，其氨基酸序列如SEQ ID NO:1所示，来源于红珊瑚弧菌（Vibrio coralliirubri）。
编码上述褐藻胶裂解酶ALVC-03的基因，其核苷酸序列如SEQ ID NO:2所示。
所述褐藻胶裂解酶ALVC-03在制备马尾藻水解物中的应用。
褐藻胶裂解酶ALVC-03的氨基酸序列（如SEQ ID NO:1所示）：
MFKKNMLAVALLSAVPIVSFANNGVSYPVPADKFDMHNWKITIPSDINEDGRVDEIEGVAMMSYSHSDFFHLDKDGNLVFEVQNQAITTKNSKNARSELRQMPRGADFSIDTADKGNQWALSSHPAASEYSAVGGTLEATLKVNHVSVNAKFPEKYPAHSVVVGQIHAKKHNELIKAGTGYGHGNEPLKIFYKKFPDQEMGSVFWNYERNLEKKDPNRADIAYPVWGNTWENPAEPGEAGIALGEEFSYKVEVKGTMMYLTFETERHDTVKYEIDLSKGIDELDSPTGYAEDDFYYKAGAYGQCSVSDSHPVWGPGCGGTGDFAVDKKNGDYNSVTFSALKLNGK。
编码褐藻胶裂解酶ALVC-03的基因序列（如SEQ ID NO:2所示）：
5’-ATGTTTAAAA AAAACATGCT GGCTGTTGCT CTGCTGTCTG CGGTCCCGAT CGTATCCTTCGCTAACAACG GCGTGTCTTA TCCGGTTCCG GCAGACAAGT TCGACATGCA CAATTGGAAA ATTACCATCCCGAGCGACAT CAACGAAGAC GGTCGCGTGG ACGAGATCGA AGGTGTCGCA ATGATGTCCT ACTCCCATTCTGACTTCTTC CACCTGGATA AAGATGGTAA CCTGGTTTTC GAAGTACAGA ACCAGGCAAT TACCACTAAAAACTCTAAGA ATGCGCGTTC CGAACTGCGT CAGATGCCGC GTGGCGCGGA TTTTTCTATC GACACTGCGGATAAGGGCAA CCAGTGGGCT CTGAGCTCCC ACCCGGCGGC AAGCGAATAC TCTGCCGTTG GTGGCACCCTGGAAGCAACT CTGAAAGTTA ACCATGTCTC TGTTAACGCA AAATTCCCGG AAAAATATCC GGCTCACTCCGTAGTAGTAG GCCAGATTCA CGCCAAAAAG CACAACGAAC TGATTAAAGC AGGTACTGGC TACGGCCACGGTAACGAACC GCTGAAAATC TTCTACAAAA AGTTCCCGGA CCAGGAGATG GGTTCTGTTT TTTGGAACTACGAACGTAAC CTGGAAAAAA AAGACCCGAA CCGTGCAGAC ATTGCCTACC CGGTTTGGGG TAATACCTGGGAAAACCCGG CTGAGCCGGG TGAGGCGGGC ATCGCTCTGG GCGAAGAGTT CTCCTATAAG GTAGAAGTCAAGGGTACCAT GATGTATCTG ACCTTCGAAA CTGAACGCCA CGATACTGTT AAATACGAAA TCGACCTGTCTAAAGGCATT GACGAGCTGG ACTCTCCGAC CGGTTACGCA GAAGACGATT TCTACTACAA GGCCGGTGCTTACGGTCAGT GCAGCGTCTC TGATAGCCAT CCGGTGTGGG GTCCAGGTTG CGGTGGTACC GGTGACTTCGCCGTTGATAA AAAAAATGGT GACTACAACA GCGTTACGTT CTCCGCTCTG AAACTGAACG GTAAA-3’。
本发明的马尾藻水解物，具有抗疲劳功能活性，能够缓解过量运动导致的机体疲劳，使机体耐力和力量得到有效恢复，具有成为缓解过量运动疲劳的功能食品原料的应用潜力。本发明的褐藻胶裂解酶ALVC-03，酶解马尾藻后可以得到抗疲劳效果更佳的酶解产物，这说明该褐藻胶裂解酶ALVC-03有着不同于现有的褐藻胶裂解酶的特性。
本发明使用的各种术语和短语具有本领域技术人员公知的一般含义。
附图说明
图1：ALVC-03酶的蛋白电泳图。
图2：ALVC-03酶的最适温度与最适pH示意图，其中，A、最适温度；B、最适pH。
图3：ALVC酶水解海藻酸钠产物的质谱检测结果示意图。
图4：各实验组的游泳时间与抓握力测试结果图，其中，A、游泳时间；B、抓握力。
图5：各实验组生理生化指标测试结果图，其中，A、BUN水平；B、LD水平；C、LG水平；D、Pi水平。
具体实施方式
下面结合实施例对本发明作进一步的说明。然而，本发明的范围并不限于下述实施例。本领域的专业人员能够理解，在不背离本发明的精神和范围的前提下，可以对本发明进行各种变化和修饰。
下述实施例中所涉及的仪器、试剂、材料等，若无特别说明，均为现有技术中已有的常规仪器、试剂、材料等，可通过正规商业途径获得。下述实施例中所涉及的实验方法，检测方法等，若无特别说明，均为现有技术中已有的常规实验方法，检测方法等。
实施例1 褐藻胶裂解酶ALVC-03的获得
来源于海洋的微生物因其特殊的生活环境，往往具有高效的海藻裂解酶活性。发明人从来源于海洋的微生物红珊瑚弧菌（Vibrio coralliirubri）的基因组中挖掘到一段褐藻胶裂解酶的基因，其经密码子优化后的基因序列与编码蛋白质序列分别如SEQ ID NO:2和SEQ ID NO:1所示，将其编码的褐藻胶裂解酶命名为ALVC-03，经密码子优化后的基因序列由人工全基因合成得到。
将ALVC-03基因以无缝拼接方式重组于pET-28a质粒载体，导入Escherichia coliBL21(DE3)中，得到可发酵生产褐藻胶裂解酶ALVC-03的工程菌株。将工程菌株以LB培养基培养至菌体OD600至0.6后，并使用IPTG诱导表达，表达后离心收集菌体，使用pH 7.0磷酸缓冲液重悬后超声破碎菌体，破碎液离心后得到上清液为粗酶液，粗酶液使用Ni柱纯化，使用80 mM咪唑洗脱可获得单一条带。蛋白电泳显示（如图1所示），该单一条带分子量约为38kDa，与该酶预测的分子量一致，表明ALVC-03纯化成功。
实施例2 褐藻胶裂解酶ALVC-03酶学性质与酶解产物研究
以190 μL 0.3%（w/v）的海藻酸钠溶液为底物，加入10 μL稀释后的ALVC-03纯酶酶液（50 U/mL），分别在30℃、40℃、50℃、60℃、70℃条件下和不同缓冲液组成的pH 3、4、5、6、7、8、9、10条件下进行反应，分别测定该酶的最适反应温度和最适反应pH。反应时间20 min，煮沸2 min灭活后使用DNS法测定产物中还原糖含量计算酶活性。实验结果如图2A、B所示，由实验结果可以看出，褐藻胶裂解酶ALVC-03的最适温度为40℃，最适pH为7.0。
以190 μL 0.3%（w/v）的海藻酸钠溶液为底物，加入10 μL稀释后的ALVC-03酶液（50 U/mL），在40℃、pH 7.0条件下反应5 h，将反应产物进行质谱检测，结果如图3所示，可以看出，该酶水解海藻酸钠底物，寡糖中的产物主要为褐藻二糖、褐藻三糖和褐藻四糖。
实施例3 马尾藻酶解液的制备
将马尾藻粉碎，使用去离子水按照固液比1:5配制成马尾藻悬浮液，马尾藻悬浮液使用匀浆仪匀浆10 min，按照马尾藻质量5%加入褐藻胶裂解酶ALVC-03粗酶液（实施例1制备，粗酶液酶活为30 U/mL），在40℃、pH 7.0条件下反应10 h，反应后121℃灭酶灭菌10min，于8000 r/min离心10 min，上清液为马尾藻酶解液Ⅰ。
将马尾藻粉碎，使用去离子水按照固液比1:5配制成马尾藻悬浮液，马尾藻悬浮液使用匀浆仪匀浆10 min，按照马尾藻质量5%加入褐藻胶裂解酶ScCD6粗酶液（该酶来源于公开号为CN 110387367A的中国发明专利，与本申请的申请人相同，使用全基因合成方式得到，异源表达方式同ALVC-03酶，粗酶液酶活为30 U/mL），在50℃、pH 9.0条件下反应10 h，反应液后121℃灭酶灭菌10 min，于8000 r/min离心10 min，上清液为马尾藻酶解液Ⅱ。
实施例4 马尾藻发酵液的制备
将马尾藻使用ALVC-03酶解后未离心酶解产物中加入1%葡萄糖，于121℃灭菌20min，按照1%（v/v）接种量接种使用LB培养基活化的枯草芽孢杆菌（菌株购买自中国普通微生物菌种保藏管理中心，菌株编号CGMCC 1.15792）种子培养液，于37℃条件下，发酵30 h，发酵完毕发酵液于121℃灭菌20 min，于8000 r/min离心10min，上清液为马尾藻发酵液Ⅰ。
将马尾藻使用ScCD6酶解后未离心酶解产物中加入1%葡萄糖，于121℃灭菌20min，按照1%（v/v）接种量接种使用LB培养基活化的枯草芽孢杆菌（菌株购买自中国普通微生物菌种保藏管理中心，菌株编号CGMCC 1.15792）种子培养液，于37℃条件下，发酵30 h，发酵完毕发酵液于121℃灭菌20 min，于8000 r/min离心10min，上清液为马尾藻发酵液Ⅱ。
实验1 马尾藻水解物小鼠实验训练
取120只雄性ICR小鼠随机分为6组，每组20只，1组：一般运动组，灌胃生理盐水；2组：过量游泳训练组，灌胃生理盐水；3组：过量游泳训练组，灌胃马尾藻酶解液Ⅰ；4组：过量游泳训练组，灌胃马尾藻发酵液Ⅰ；5组：过量游泳训练组，灌胃马尾藻酶解液Ⅱ；6组：过量游泳训练组，灌胃马尾藻发酵液Ⅱ。各组灌胃剂量均为10 mL/kg体重，每日灌胃一次，持续7周。所有实验组第1周进行无负载游泳训练，第2,3,4实验组于第2-6周进行负载游泳训练，期间第1组继续进行无负载游泳训练。
游泳训练规则：水深20 cm，水温25±2℃，第1天训练30 min，第二天训练45 min，其余训练日训练60 min，一周训练6天。第2周至第6周，过量游泳训练组尾部固定重物，每周增加体重2%的质量，最终达到10%体重。每次训练时，小鼠沉入水底超过3 s，游泳训练即结束，此时立刻将小鼠移出水中。
实验2 马尾藻水解物小鼠实验测试
第7周进行小鼠承重游泳测试和抓握力测试。
承重游泳测试：所有实验组第43日进行负载游泳测试，尾部重物质量为小鼠体重10%，方法与上述游泳规则相同，沉水时间增加至8 s视为游泳结束。
抓握力测试：所有实验组第46日进行抓握力测试，提起小鼠尾部，其前爪抓握网栅，轻柔提拽小鼠，测定小鼠被提起过程的最大抓握力，每只小鼠测定3次，取平均值。
生理生化指标测定：所有实验组于第49日进行15 min负载游泳，游泳后腹腔注射戊巴比妥钠麻醉处死，取血液、肝脏和腓肌肠进行各项生化指标测定。
马尾藻水解物小鼠实验结果：
各实验组小鼠的承重游泳测试和抓握力测试结果如图4所示。承重游泳测试（图4A）显示，实验2组的游泳时间最短，表明过量运动导致的疲劳会明显降低小鼠的耐力；实验3组与实验1组游泳时间无显著性差异，但显著长于实验2组，表明灌胃马尾藻酶解液Ⅰ，可以一定程度上缓解过量运动疲劳导致的耐力下降；实验4组的游泳时间最长，说明马尾藻发酵液Ⅰ灌胃效果明显，可显著改善过量运动疲劳导致的耐力下降；实验5组游泳时间长于实验2组游泳时间，但短于实验组3游泳时间，说明马尾藻酶解液Ⅱ有助于缓解过量运动疲劳导致的耐力下降，但效果明显弱于马尾藻酶解液Ⅰ；实验6组游泳时间长于实验2组和实验5组，与实验1组和实验3组相当，无显著性差异，短于实验4组，说明马尾藻发酵液Ⅱ缓解过量运动疲劳导致的耐力下降效果强于马尾藻酶解液Ⅱ，但明显弱于马尾藻发酵液Ⅰ。
抓握力测试（图4B）结果分析显示，其中实验4组抓握力最高，实验2组抓握力最低，实验5组抓握力与实验2组无显著性差异，实验1组、实验3组和实验5组抓握力无显著性差异，说明马尾藻发酵液Ⅰ、马尾藻酶解液Ⅰ和马尾藻发酵液Ⅱ，都可有效缓解过量运动疲劳导致的力量下降，其中马尾藻发酵液Ⅰ的改善作用最明显。
各实验组小鼠的生理生化指标测试结果如图5所示。血尿素氮（BUN,图5A）分析结果显示，实验2组的血尿氮素含量最高，实验5组的血尿氮素含量最高，实验1组、实验3组和实验6组的血尿氮素相近，实验4组的血尿氮素最低。血尿胆素反映机体的代谢能力和运动耐力，机体疲劳度增加时，往往会导致血尿胆素含量升高，可以看出马尾藻酶解液Ⅰ、马尾藻酶解液Ⅱ、马尾藻发酵液Ⅰ和马尾藻发酵液Ⅱ都可以不同程度上缓解机体疲劳导致的血尿氮素含量升高，其中马尾藻发酵液Ⅰ的缓解效果最明显。
乳酸（LD，图5B）分析结果显示，实验2组和实验5组血液中乳酸含量最高，实验1组、实验3组和实验6组血液中乳酸含量相近，实验4组血液中乳酸含量最低。在剧烈运动时，肌肉细胞的有氧呼吸转变为糖酵解无氧呼吸，肌肉中的糖原会快速消耗产生大量的乳酸。乳酸在肌肉和血液中的积累会导致肌肉乏力的现象产生，可以看出马尾藻酶解液Ⅰ、马尾藻发酵液Ⅰ和马尾藻发酵液Ⅱ都可以在不同程度上降低肌肉细胞乳酸的形成，其中马尾藻发酵液Ⅰ的效果最明显。
肝糖原（LG，图5C）分析结果显示，实验2组的肝糖原含量最低，实验5组的肝糖原含量较实验2组稍高，实验1组、实验3组和实验6组的肝糖原含量相近，实验4组的肝糖原含量最高，肝糖原最要最为能量储备，可以在运动中持续向机体提供能量。可以看出，可以看出马尾藻酶解液Ⅰ、马尾藻酶解液Ⅱ、马尾藻发酵液Ⅰ和马尾藻发酵液Ⅱ都可以不同程度增加机体肝糖原含量，其中马尾藻发酵液Ⅰ的效果最好。
无机磷酸盐（Pi，图5D）分析结果显示，实验2组和实验5组的腓肠肌细胞中的无机磷酸盐含量最高，实验1组、实验3组和实验6组的无机磷酸盐含量相近，实验4组的无机磷酸盐含量最低。细胞中无机磷酸盐的存在会直接作用于肌肉横桥、降低肌纤维的敏感度，从而导致肌肉力量的降低。可以看出，马尾藻酶解液Ⅰ、马尾藻发酵液Ⅰ和马尾藻发酵液Ⅱ都可以不同程度上减低腓肠肌细胞中无机磷酸研的含量。
综合以上实验结果可以看出，使用ALVC-03酶酶解处理后，再进行枯草芽孢杆菌发酵制得的马尾藻发酵液可以较为明显地提升小鼠过量运动疲劳导致的耐力下降和力量下降。马尾藻经ALVC-03酶酶解后所得的酶解产物，在提升小鼠过量运动疲劳导致的耐力下降和力量下降方面，明显优于褐藻胶裂解酶ScCD6酶解后所得的酶解产物，可能是由于褐藻胶裂解酶ALVC-03酶解后产物褐藻寡糖的聚合度不同于褐藻胶裂解酶ScCD6酶酶解后产物褐藻寡糖的聚合度，不同聚合度褐藻寡糖在提升小鼠过量运动导致的耐力下降和力量下降方面的功能活性不同，导致两种制备工艺得到的产物的功能活性的不同。马尾藻发酵液制备过程绿色环保，无有害化学物质添加，且原料为纯天然海藻成分，在安全性和功能性上都具有作为抗运动疲劳功能食品原料使用的应用潜力。
给本领域技术人员提供上述实施例，以完全公开和描述如何实施和使用所主张的实施方案，而不是用于限制本文公开的范围。对于本领域技术人员而言显而易见的修饰将在所附权利要求的范围内。
序列表
<110> 中国海洋大学
<120> 具有抗疲劳功能活性的马尾藻水解物及其制备方法与应用
<141> 2021-10-15
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Claims (8)
1.一种制备马尾藻水解物的方法，其特征在于：将马尾藻粉碎，加水配制马尾藻悬浮液；加入褐藻胶裂解酶ALVC-03或含褐藻胶裂解酶ALVC-03的酶液，酶解；酶解后加入葡萄糖，高压灭菌，接种枯草芽孢杆菌，发酵；发酵结束后高压灭菌，离心，上清液即为具有抗疲劳功能活性的马尾藻水解物；所述褐藻胶裂解酶ALVC-03，其氨基酸序列如SEQ ID NO:1所示。
2.根据权利要求1所述的制备马尾藻水解物的方法，其特征在于：所述枯草芽孢杆菌，选自保藏编号为CGMCC 1.15792的枯草芽孢杆菌。
3.根据权利要求1所述的制备马尾藻水解物的方法，其特征在于：所述马尾藻悬浮液，马尾藻与水的质量体积比为1:3～8；
所述褐藻胶裂解酶ALVC-03的质量为马尾藻质量的3%～8%；
所述酶解的条件为：温度35～45℃，pH 6.5～7.5，时间8～12小时。
4.根据权利要求1所述的制备马尾藻水解物的方法，其特征在于：所述葡萄糖的添加量为酶解液体积的0.5%～2%；
所述发酵的条件为：35～40℃发酵25～35小时。
5.根据权利要求1所述的制备马尾藻水解物的方法，其特征在于：所述马尾藻悬浮液，马尾藻与水的质量体积比为1:5；
所述褐藻胶裂解酶ALVC-03的质量为马尾藻质量的5%；
所述酶解的条件为：温度40℃，pH 7.0，时间10小时；
所述葡萄糖的添加量为酶解液体积的1%；
所述高压灭菌的条件为：121℃灭菌20 min；
所述发酵的条件为：37℃发酵30小时；
所述离心的条件为：8000 r/min离心10 min。
6.利用权利要求1～5中任一项所述的方法制备得到的具有抗疲劳功能活性的马尾藻水解物。
7.权利要求6所述的具有抗疲劳功能活性的马尾藻水解物在制备缓解运动疲劳的功能性食品或饮料中的应用。
8.褐藻胶裂解酶ALVC-03在制备马尾藻水解物中的应用，所述褐藻胶裂解酶ALVC-03的氨基酸序列如SEQ ID NO:1所示。
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040088416A (ko) * | 2004-09-15 | 2004-10-16 | 윤종선 | 신규한 해양 미생물 잔토모나스 에스피. 비에이치-1을이용한 알긴산 분해효소의 제조방법 |
| CN104293754A (zh) * | 2014-09-12 | 2015-01-21 | 山东大学 | 一种内切型褐藻胶裂解酶及其编码基因与应用 |
| WO2015104723A1 (en) * | 2014-01-10 | 2015-07-16 | Matis Ohf. | Thermostable alginate degrading enzymes and their methods of use |
| CN106509895A (zh) * | 2016-11-08 | 2017-03-22 | 全椒先奇医药科技有限公司 | 一种益生菌保健品及其制备方法 |
| CN107400589A (zh) * | 2017-09-21 | 2017-11-28 | 青岛金海宝生物科技发展有限公司 | 一种富含褐藻寡糖的葡萄酒 |
| KR101918239B1 (ko) * | 2017-06-20 | 2018-11-13 | 재단법인 전남생물산업진흥원 | 괭생이 모자반 가수분해물을 유효성분으로 함유하는 고혈압 예방 또는 치료용 약학적 조성물 |
| CN110387367A (zh) * | 2019-09-03 | 2019-10-29 | 中国海洋大学 | 一种新型褐藻胶裂解酶 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012142326A1 (en) * | 2011-04-12 | 2012-10-18 | Bio Architecture Lab, Inc. | A method of producing 5-hydroxypyridine-2-carboxylic acid from alginate |
-
2021
- 2021-10-25 CN CN202111238077.9A patent/CN113662181B/zh active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040088416A (ko) * | 2004-09-15 | 2004-10-16 | 윤종선 | 신규한 해양 미생물 잔토모나스 에스피. 비에이치-1을이용한 알긴산 분해효소의 제조방법 |
| WO2015104723A1 (en) * | 2014-01-10 | 2015-07-16 | Matis Ohf. | Thermostable alginate degrading enzymes and their methods of use |
| CN104293754A (zh) * | 2014-09-12 | 2015-01-21 | 山东大学 | 一种内切型褐藻胶裂解酶及其编码基因与应用 |
| CN106509895A (zh) * | 2016-11-08 | 2017-03-22 | 全椒先奇医药科技有限公司 | 一种益生菌保健品及其制备方法 |
| KR101918239B1 (ko) * | 2017-06-20 | 2018-11-13 | 재단법인 전남생물산업진흥원 | 괭생이 모자반 가수분해물을 유효성분으로 함유하는 고혈압 예방 또는 치료용 약학적 조성물 |
| CN107400589A (zh) * | 2017-09-21 | 2017-11-28 | 青岛金海宝生物科技发展有限公司 | 一种富含褐藻寡糖的葡萄酒 |
| CN110387367A (zh) * | 2019-09-03 | 2019-10-29 | 中国海洋大学 | 一种新型褐藻胶裂解酶 |
Non-Patent Citations (3)
| Title |
|---|
| "Isolation of a novel alginate lyase-producing Bacillus litoralis strain and its potential to ferment Sargassum horneri for biofertilizer";Mingpeng Wang et al;《MicrobiologyOpen》;20161231;第5卷;第1038-1049页 * |
| "WP_04607052.1 MULTISPECIES:polysaccharide lyase family 7 protein[vibrio]";NCBI SPARCLE;《GenPept》;20210309;第1页 * |
| "铜藻发酵提取物的制备及其抗疲劳活性研究";邢义高;《中国优秀博硕士学位论文全文数据库(硕士) 工程科技Ⅰ辑》;20210215(第02期);第B024-1108页 * |
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