CN113667661A - 一种β-葡萄糖苷酶及其在制备葡萄糖和昆布寡糖中的应用 - Google Patents
一种β-葡萄糖苷酶及其在制备葡萄糖和昆布寡糖中的应用 Download PDFInfo
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- CN113667661A CN113667661A CN202110846319.6A CN202110846319A CN113667661A CN 113667661 A CN113667661 A CN 113667661A CN 202110846319 A CN202110846319 A CN 202110846319A CN 113667661 A CN113667661 A CN 113667661A
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- Prior art keywords
- glucosidase
- beta
- laminarin
- bglh
- gly
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Abstract
本发明涉及一种β‑葡萄糖苷酶及其在制备葡萄糖和昆布寡糖中的应用。所述β‑葡萄糖苷酶氨基酸序列如SEQ ID NO.1所示。本发明的β‑葡萄糖苷酶为结构与功能新颖的同时具有葡聚糖酶和转糖苷酶活性的水解酶,其氨基酸序列与已有性质报道的β‑葡萄糖苷酶序列相似度仅为78.96%,是一个新酶。本发明所述的β‑葡萄糖苷酶产量可达406.1U/mL,最适反应温度为40℃,最适反应PH为6。本发明所述的β‑葡萄糖苷酶可以同时表现出针对β‑葡聚糖的葡聚糖酶活性生产葡萄糖和针对昆布多糖的转糖苷酶活性生产昆布寡糖。
Description
技术领域
本发明涉及一种β-葡萄糖苷酶及其在制备葡萄糖和昆布寡糖中的应用,属于生物技术领域。
背景技术
β-葡萄糖苷酶(β-D-Glucosidase,EC3.2.1.21),又称β-D-葡萄糖苷葡萄糖水解酶,别名龙胆二糖酶、纤维二糖酶(cellobias,CB或β-G)和苦杏仁苷酶。它属于纤维素酶类,是纤维素分解酶系中的重要组成成分,能够水解结合于末端非还原性的β-D-葡萄糖键,同时释放出β-D-葡萄糖和相应的配基。木质纤维素生物质由于丰富的资源及低廉的成本,长期以来被认为是用于生产可发酵糖(例如葡萄糖)的可再生生物质。然而,其具有高含量的木质素及非常复杂的结构,且因此需要高强度的预处理及酶促糖基化,从而使其难以使用。近来,海藻作为替代木质纤维素生物质的生物质受到广泛关注,且在高碳水化合物含量、不需要耕地以及由于几乎没有木质素含量而具有简单结构方面是有利的。
在海洋藻类中,全世界每年收获约70百万吨褐藻,褐藻的主要碳水化合物成分是海藻酸及葡聚糖,且葡聚糖主要是昆布多糖。昆布多糖在其骨架中具有β-1,3键,且在其分支中具有β-1,6键。在褐藻中,昆布属(Laminarina)、海带属 (Saccharina)及岩藻属(Fucusspp.)具有30%到80%干重的高昆布多糖含量。由于昆布多糖是由葡萄糖组成的多糖,因此昆布多糖是用于生产可发酵糖的非常理想的生物质。此外,已知从昆布多糖生产的昆布寡糖具有多种生理活性,且因此可用作功能材料。
为使用木质纤维素生物质生产可发酵糖,需要高强度的预处理,且其需要至少三种酶的组合,例如内切葡聚糖酶、外切葡聚糖酶及β-糖苷酶。这种预处理工艺使用高热量,且因此需要大量能量,并且在酶促糖基化工艺中使用的酶的成本占整个生物工艺的很大一部分。
因此,需要一种通过使用最少数量的酶以降低的生产成本高效地水解昆布多糖来生产作为发酵糖的葡萄糖及作为功能材料的昆布寡糖的工艺。
发明内容
本发明针对现有技术的不足,提供了一种新型β-葡萄糖苷酶BglH及其制备方法。本发明的β-葡萄糖苷酶BglH为结构与功能新颖的纤维素酶,其氨基酸序列与已有性质报道的β-葡萄糖苷酶序列相似度仅为78.96%,是一个新酶。本发明所述一种新型β-葡萄糖苷酶BglH最适反应温度为40℃,最适反应pH为6;其同时具有外切性葡聚糖酶活性及转糖苷酶活性,是用于制备葡萄糖和昆布寡糖的理想选择。
一方面,本发明提供一种新型β-葡萄糖苷酶BglH,其氨基酸序列如SEQ ID NO.1所示。
SEQ ID NO.1:
MVKSYVMKHASPVGRRAFPIIYYKFGMRSNVRESVSSIVLAVIQYFAKGL FRALPEYQRSLYILGVFCAHKESDKFMSIHMFPSDFKWGVATAAYQIEGAYNE DGRGMSIWDTFAHTPGKVKNGDNGNVACDSYHRVEEDVQLLKDLGVKVYR FSISWPRVLPQGTGEVNRAGLDYYHRLVDELLANGIEPFCTLYHWDLPQALQ DQGGWGSRITIDAFAEYAELMFKELGGKIKQWITFNEPWCMAFLLSNYLGVH APGNKDLQLAIDVSHHLLVAHGRAVTLFRELGISGEIGIAPNTSWAVPYRRTKE DMEACLRVNGWSGDWYLDPIYFGEYPKFMLDWYENLGYKPPIVDGDMELIH QPIDFIGINYYTSSMNRYNPGEAGGMLSSEAISMGAPKTDIGWEIYAEGLYDLL RYTADKYGNPTLYITCYNDGLSLDGRIHDQRRIDYLAMHLENGAIQASRAIED GINLKGYMEWSLMDNFEWAEGYGMRFGLVHVDYDTLVRTPKDSFYWYK。
另一方面,本发明还提供一种新型β-葡萄糖苷酶BglH对应的核酸序列,如SEQ IDNO.2所示。
SEQ ID NO.2:
atggtgaaaagctatgtgatgaaacatgcgagcccggtgggccgccgcgcgtttccgattatttattataaatttggc atgcgcagcaacgtgcgcgaaagcgtgagcagcattgtgctggcggtgattcagtattttgcgaaaggcctgtttcgcgcg ctgccggaatatcagcgcagcctgtatattctgggcgtgttttgcgcgcataaagaaagcgataaatttatgagcattcatatg tttccgagcgattttaaatggggcgtggcgaccgcggcgtatcagattgaaggcgcgtataacgaagatggccgcggcat gagcatttgggatacctttgcgcataccccgggcaaagtgaaaaacggcgataacggcaacgtggcgtgcgatagctatc atcgcgtggaagaagatgtgcagctgctgaaagatctgggcgtgaaagtgtatcgctttagcattagctggccgcgcgtgc tgccgcagggcaccggcgaagtgaaccgcgcgggcctggattattatcatcgcctggtggatgaactgctggcgaacgg cattgaaccgttttgcaccctgtatcattgggatctgccgcaggcgctgcaggatcagggcggctggggcagccgcattac cattgatgcgtttgcggaatatgcggaactgatgtttaaagaactgggcggcaaaattaaacagtggattacctttaacgaac cgtggtgcatggcgtttctgctgagcaactatctgggcgtgcatgcgccgggcaacaaagatctgcagctggcgattgatg tgagccatcatctgctggtggcgcatggccgcgcggtgaccctgtttcgcgaactgggcattagcggcgaaattggcattg cgccgaacaccagctgggcggtgccgtatcgccgcaccaaagaagatatggaagcgtgcctgcgcgtgaacggctgga gcggcgattggtatctggatccgatttattttggcgaatatccgaaatttatgctggattggtatgaaaacctgggctataaacc gccgattgtggatggcgatatggaactgattcatcagccgattgattttattggcattaactattataccagcagcatgaaccg ctataacccgggcgaagcgggcggcatgctgagcagcgaagcgattagcatgggcgcgccgaaaaccgatattggctg ggaaatttatgcggaaggcctgtatgatctgctgcgctataccgcggataaatatggcaacccgaccctgtatattacctgct ataacgatggcctgagcctggatggccgcattcatgatcagcgccgcattgattatctggcgatgcatctggaaaacggcg cgattcaggcgagccgcgcgattgaagatggcattaacctgaaaggctatatggaatggagcctgatggataactttgaat gggcggaaggctatggcatgcgctttggcctggtgcatgtggattatgataccctggtgcgcaccccgaaagatagctttta ttggtataaa。
另一方面,本发明还提供一种β-葡萄糖苷酶BglH的制备与纯化方法。
另一方面,本发明还提供了所述β-葡萄糖苷酶BglH在制备葡萄糖和昆布寡糖的应用。
另一方面,一种降解β-葡聚糖和昆布多糖的方法,所选用的β-葡萄糖苷酶为BglH。
优选:所述降解条件中反应温度为0~70℃。最适反应温度为40℃。
优选:所述降解条件中反应pH为5.5~10.5。最适反应pH为6。
有益效果:
1.本发明的β-葡萄糖苷酶BglH为结构与功能新颖的纤维素酶,其氨基酸序列与已有性质报道的β-葡萄糖苷酶序列相似度仅为78.96%。
2.本发明提供了一种制备β-葡萄糖苷酶BglH的方法,即利用基因工程的技术方法,将BglH的基因序列异源重组表达到大肠杆菌,经发酵后,发酵液酶活力高达406.1U/mL,具有工业化生产的潜质。该酶纯化方法简单,可利用镍柱对其进行一步亲和纯化。
3.本发明的β-葡萄糖苷酶BglH具有优良的理化性质,该酶最适反应温度和 pH分别为40℃和6,且其同时具有外切性葡聚糖酶活性及转糖苷酶活性。本发明所述的β-葡萄糖苷酶BglH具有良好的工业化应用前景。
附图说明
图1为本发明β-葡萄糖苷酶BglH蛋白质分离纯化图(M,蛋白质标准品; 1,纯化所得β-葡萄糖苷酶BglH);
图2为本发明β-葡萄糖苷酶BglH的温度和pH适应性分析(A,β-葡萄糖苷酶BglH的最适反应温度;B,β-葡萄糖苷酶BglH的最适反应pH;
图3为本发明β-葡萄糖苷酶BglH的热稳定性分析;
图4为本发明β-葡萄糖苷酶BglH在高浓度条件下的最终酶解产物的高效液相色谱图。
图5为本发明β-葡萄糖苷酶BglH在低浓度条件下的最终酶解产物的高效液相色谱图。
具体实施方式
实施例1β-葡萄糖苷酶BglH的序列分析及重组表达
本发明所述β-葡萄糖苷酶BglH的产酶基因BglH来源于海洋细菌Baciliiussp.HY29,包含有1551个碱基序列,编码517个氨基酸序列。利用National Center forBiotechnology Information(NCBI)中的保守结构域分析Conserved domain (CDD)和多重序列比对Basic Local Alignment Search Tool(Blast)发现,该序列包含有一段BGL的β-葡萄糖苷酶保守区。已经报道的β-葡萄糖苷酶中,与BglH 氨基酸序列相似度最高的为β-葡萄糖苷酶(bglA)(Genbank M96979.1),两者之间的氨基酸序列相似度(Identity)为78.96%。本发明所述β-葡萄糖苷酶 BglH序列新颖。
本发明还提供一种生产葡萄糖或昆布寡糖的方法,所述方法包括使具有氨基酸序列SEQ ID NO.1的β-葡萄糖苷酶与昆布多糖或昆布二糖反应,从而生产葡萄糖或昆布寡糖。
此外,缓冲液中的β-葡萄糖苷酶的最佳pH可根据缓冲液的类型而变化,但β-葡萄糖苷酶在约pH 5到约pH 7.5时表现出80%或大于80%的酶活性,且特别是在pH为6时表现出最高活性。此外,β-葡萄糖苷酶在pH为6及10℃到 40℃下表现出80%或大于80%的酶活性。然而,在50℃或高于50℃下,酶活性急剧降低,且因此,由于β-葡萄糖苷酶即使在室温下也可引起充分的酶促反应,因此存在的优点是生产工艺可经济地执行,而不需要升高温度的能量消耗。
将β-葡萄糖苷酶BglH的产酶序列以限制性内切酶Nco I和Xho I为酶切位点,设计重组引物如下(下划线为限制性内切酶位点,斜体为限制性内切酶保护碱基):
正向引物:SEQ ID NO.3:PBglH-F:
5’-CATGCCATGGAAGTTGTCTTGTATCGCT-3’(Nco I)
反向引物:SEQ ID NO.4:PBglH-R:
5’-CCGCTCGAGCTTGATTTCGTATGGGTCA-3’(Xho I)
PCR扩增条件为:94℃预变性3min;94℃变性30秒,55℃退火30秒,72℃延伸1min,共30个循环;72℃延伸5min;4℃稳定15min。PCR反应所用DNA 聚合酶为Primerstar HS购自大连宝生物公司。
PCR产物用限制性内切酶Nco I和Xho I进行双酶切,通过琼脂糖凝胶电泳回收酶切后的PCR产物。pET28a(+)质粒DNA(美国Invitrogen公司),同样用限制性内切酶Nco I和Xho I进行双酶切,进行琼脂糖凝胶电泳并回收酶切后的产物片段。酶切所用酶和底物反应体系(温度、时间、DNA用量等)均参照大连宝生物提供的产品说明操作。
双酶切处理的PCR产物和pET-28a(+)质粒载体参照DNA连接酶(大连宝生物公司)说明书进行连接反应;连接产物转化至大肠杆菌DH5α菌株(美国 Invitrogen公司),涂布在Luria-Bertani(LB)培养基固体平板上(含有50μg/mL 卡那霉素),37℃温箱中培养12-16小时后,挑取单克隆;将单克隆转接至LB 液体培养基中(含有50μg/mL卡那霉素),转速为160rpm的37℃摇床中培养过夜。将单克隆进行序列测定,选择阳性克隆,并将其命名为pET28a-BglH。重组质粒转化至大肠杆菌BL21(DE3)(购自大连宝生物公司),将重组大肠菌株命名为BL21(DE3)/pET28a-BglH,保存在-80℃备用。
实施例2β-葡萄糖苷酶BglH的制备及纯化方法
将重组菌株BL21(DE3)/pET28a-BglH在100mL的LB液体培养基中(50 μg/mL卡那霉素),在37℃摇床中160rpm震荡培养至OD600=0.6,加入终浓度为0.1mM的诱导剂异丙基-β-D-硫代半乳糖苷(IPTG),在20℃诱导24h。β- 葡萄糖苷酶活性测定方法为:50μL酶液加入450μL 0.1%(w/v)纤维二糖,在40℃下反应15min。将混合物以10,000rpm离心10min,在OD420下检测其吸光度。酶活力定义为1U为每min产生1μM葡萄糖所需要的酶量。经检测,发酵液中β-葡萄糖苷酶活力可达406.1U/mL。
发酵停止后,12000rpm离心10min,弃上清液后收集菌体。将菌体重悬于 20mM磷酸盐缓冲液中,使用超声细胞破碎仪破碎菌体(全程操作在冰上,保持低温)。最后,将细菌裂解物离心并收集上清液,使用Akta150 FPLC纯化系统进行纯化。将收集的上清液上样于经过预平衡的5mL镍离子亲和层析柱上,上样流速为5mL/min。洗涤缓冲液(500mM NaCl,20mM磷酸缓冲液,PH7.6) 用于去除杂质蛋白,使用洗脱缓冲液(500mM咪唑,500mM NaCl,20mM磷酸缓冲液,PH7.6)用于活性成分。将活性成分透析去除咪唑,分装储存在-20℃备用。纯化所得β-葡萄糖苷酶进行聚丙烯酰胺凝胶电泳(SDS-PAGE),如图1 所示,纯化所得β-葡萄糖苷酶BglH的分子量为59kDa,与序列分析中预测的蛋白大小一致。
实施例3β-葡萄糖苷酶BglH的最适温度和pH测定
将实施例2中纯化所得β-葡萄糖苷酶BglH在不同条件下进行酶活力测定,检测不同温度和pH对酶活力的影响。在不同温度(0-60℃)下反应15min,检测不同反应温度对酶活力的影响,以最高酶活力为100%,计算不同温度下BglH 的相对酶活力。如图2A所示,β-葡萄糖苷酶BglH的最适反应温度为40℃。在 20℃下,保持约84.5%的酶活性。此外,在非常低的温度(即5℃到10℃)下,表现出高于60.3%的酶活性。然而,在高于50℃下,酶活性急剧降低。这表明即使在室温下也可充分执行酶促反应,且因此可在不施加升高温度的能量的情况下执行经济的工艺。此外,仅通过简单的热处理便可使酶失活。
将实施例2中纯化所得β-葡萄糖苷酶BglH与使20mM乙酸钠(pH 4.0到pH 6.0)、20mM磷酸钠(pH 6.0到pH 7.0)及20mM Tris-HCl-NaCl(pH 7.0到pH 9.0) 缓冲液(含有0.1%(w/v)的纤维二糖)在40℃下反应15分钟。在最适温度下检测活性,酶活性最高值为100%。如图2B所示,β-葡萄糖苷酶BglH的最适反应pH 为6.0。
实施例4β-葡萄糖苷酶BglH的热稳定性测定
将实施例2中纯化所得β-葡萄糖苷酶在不同稳定下反应1小时后,进行酶活性的检测。如图3所示,BglH在40℃反应1小时后,保持83%的酶活性。
实施例5β-葡萄糖苷酶BglH酶解产物HPLC分析
将实施例2中纯化所得β-葡萄糖苷酶BglH纯酶与昆布多糖在40℃下进行反应,检测其酶解产物。将100μL纯化的BglH和900μL昆布多糖底物(5mg/mL) 在20mM磷酸钠缓冲液(PH 6.0)中于40℃下孵育12小时,并通过煮沸10分钟使酶失活,然后使用高效液相色谱(High Performance Liquid Chromatography, HPLC)。具体为:使用配备有凝胶渗透及配体交换柱(KS-802;昭和电工公司 (Shodex))的安捷伦(Agilent)1100HPLC及折射率检测器(refractive index detector)执行了HPLC分析,且分析条件是流速为0.5毫升/分钟,柱温为80℃,且无菌水为流动相。对于TLC分析,将1μl反应产物装载到硅胶60板(默克公司(Merck))上,且然后使用正丁醇:乙酸:水(体积比为3:2:2)的混合溶剂进行了显影,并用10%(v/v)的硫酸进行了处理以实现可视化。如图4所示,β-葡萄糖苷酶BglH 酶解主产物为葡萄糖。
此外,测定了BglH在低浓度条件下的酶解产物,将10μL纯化的BglH和 990μL昆布多糖底物(5mg/mL)在20mM磷酸钠缓冲液(PH 6.0)中于40℃下孵育12小时后。结果如图5,生产了具有低聚合度的寡糖,例如DP2、DP3、 DP4、DP5和DP6。
从上述结果可以看出,本发明所得β-葡萄糖苷酶BglH具有工业适用性,可应用于通过酶促反应生产葡萄糖及昆布寡糖的领域。
序列表
<110> 青岛大学
<120> 一种β-葡萄糖苷酶及其在制备葡萄糖和昆布寡糖中的应用
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Claims (8)
1.一种β-葡萄糖苷酶,其氨基酸序列如SEQ ID NO.1所示。
2.如权利要求1所述的β-葡萄糖苷酶所对应的核苷酸序列,所述核苷酸序列如SEQ IDNO.2所示。
3.如权利要求1所述的β-葡萄糖苷酶的制备与纯化方法。
4.如权利要求1所述的β-葡萄糖苷酶在制备葡萄糖和昆布寡糖的应用。
5.一种制备葡萄糖和昆布寡糖的方法,其特征是,所选用的β-葡萄糖苷酶为权利要求1所述的β-葡萄糖苷酶。
6.如权利要求5所述的方法,其特征是,降解条件中反应温度为0~70℃,最适反应温度为40℃。
7.如权利要求5所述的方法,其特征是,降解条件中反应pH为5.5~10.5,最适反应pH为6。
8.如权利要求5所述的方法,其特征是,所述的β-葡萄糖苷酶同时具有外切性葡聚糖酶活性及转糖苷酶活性。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114196655A (zh) * | 2021-12-21 | 2022-03-18 | 中国海洋大学 | 耐热性昆布多糖降解酶OUC-SaLam66及其应用 |
CN114410611A (zh) * | 2021-12-21 | 2022-04-29 | 中国海洋大学 | 昆布多糖降解酶OUC-BsLam26及其应用 |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102477417A (zh) * | 2010-11-24 | 2012-05-30 | 中国农业大学 | 一种β-葡萄糖苷酶及其编码基因与应用 |
CN102925469A (zh) * | 2012-09-18 | 2013-02-13 | 湖南农业大学 | 一种编码β-葡萄糖苷酶的基因及应用 |
US20160362669A1 (en) * | 2014-05-29 | 2016-12-15 | Guangzhou Institute Of Energy Conversion, Chinese Academy Of Sciences | THERMOPHILIC ETHANOL-RESISTANT ß-GLUCOSIDASE AND ENCODING GENE AND APPLICATION THEREOF |
CN111836900A (zh) * | 2018-03-12 | 2020-10-27 | 高丽大学校产学协力团 | 从海藻生产葡萄糖及昆布多糖寡糖的新型β-葡萄糖苷酶 |
CN112292450A (zh) * | 2017-09-25 | 2021-01-29 | 味之素株式会社 | 蛋白质的制造方法和二糖的制造方法 |
CN112351990A (zh) * | 2018-04-05 | 2021-02-09 | 合同酒精株式会社 | 来自饲料类芽孢杆菌(Paenibacillus pabuli)的能够制造低聚半乳糖的酶及制造低聚半乳糖的方法 |
CN112410321A (zh) * | 2020-11-26 | 2021-02-26 | 昆明理工大学 | 一种β-葡萄糖苷酶Ttbgl3及其应用 |
-
2021
- 2021-07-26 CN CN202110846319.6A patent/CN113667661B/zh active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102477417A (zh) * | 2010-11-24 | 2012-05-30 | 中国农业大学 | 一种β-葡萄糖苷酶及其编码基因与应用 |
CN102925469A (zh) * | 2012-09-18 | 2013-02-13 | 湖南农业大学 | 一种编码β-葡萄糖苷酶的基因及应用 |
US20160362669A1 (en) * | 2014-05-29 | 2016-12-15 | Guangzhou Institute Of Energy Conversion, Chinese Academy Of Sciences | THERMOPHILIC ETHANOL-RESISTANT ß-GLUCOSIDASE AND ENCODING GENE AND APPLICATION THEREOF |
CN112292450A (zh) * | 2017-09-25 | 2021-01-29 | 味之素株式会社 | 蛋白质的制造方法和二糖的制造方法 |
CN111836900A (zh) * | 2018-03-12 | 2020-10-27 | 高丽大学校产学协力团 | 从海藻生产葡萄糖及昆布多糖寡糖的新型β-葡萄糖苷酶 |
US20210040522A1 (en) * | 2018-03-12 | 2021-02-11 | Korea University Research And Business Foundation | Novel beta-glucosidase for producing glucose and laminarioligosaccharide from sea weed |
CN112351990A (zh) * | 2018-04-05 | 2021-02-09 | 合同酒精株式会社 | 来自饲料类芽孢杆菌(Paenibacillus pabuli)的能够制造低聚半乳糖的酶及制造低聚半乳糖的方法 |
CN112410321A (zh) * | 2020-11-26 | 2021-02-26 | 昆明理工大学 | 一种β-葡萄糖苷酶Ttbgl3及其应用 |
Non-Patent Citations (2)
Title |
---|
章文明: "灰盖鬼伞菌柄细胞壁伸长特性及细胞壁蛋白的功能研究", 博士电子期刊库, no. 12, pages 1 - 140 * |
郑芳芳;王金佩;林宇;王子龙;韦宇拓;黄日波;杜丽琴;: "链霉菌GXT6 β-葡萄糖苷酶的酶学性质及葡萄糖耐受性分子改造", 微生物学报, no. 10, pages 1839 - 1852 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114196655A (zh) * | 2021-12-21 | 2022-03-18 | 中国海洋大学 | 耐热性昆布多糖降解酶OUC-SaLam66及其应用 |
CN114410611A (zh) * | 2021-12-21 | 2022-04-29 | 中国海洋大学 | 昆布多糖降解酶OUC-BsLam26及其应用 |
CN114196655B (zh) * | 2021-12-21 | 2022-11-15 | 中国海洋大学 | 耐热性昆布多糖降解酶OUC-SaLam66及其应用 |
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