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琼胶寡糖鉴定方法的研究进展 被引量:6
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作者 张伟宾 刘丹 +2 位作者 许竞男 于文功 路新枝 《中国海洋药物》 CAS CSCD 2017年第6期96-102,共7页
琼胶的主要成分琼脂糖,是由β-D-半乳吡喃糖和3,6-内醚-α-L-半乳吡喃糖通过α(1→3)和β(1→4)糖苷键交替形成的线性多糖。琼胶寡糖是琼胶多糖的降解产物,分为琼寡糖和新琼寡糖,其中琼寡糖是α-琼胶酶或酸水解形成的以3,6-内醚-α-L-... 琼胶的主要成分琼脂糖,是由β-D-半乳吡喃糖和3,6-内醚-α-L-半乳吡喃糖通过α(1→3)和β(1→4)糖苷键交替形成的线性多糖。琼胶寡糖是琼胶多糖的降解产物,分为琼寡糖和新琼寡糖,其中琼寡糖是α-琼胶酶或酸水解形成的以3,6-内醚-α-L-半乳吡喃糖为还原端、β-D-半乳吡喃糖为非还原端的寡糖产物,而β-琼胶酶降解琼胶的产物是以β-D-半乳吡喃糖为还原端的新琼寡糖。对琼胶寡糖类型的判断是琼胶酶分类鉴定的主要依据之一,本文综述了利用核磁、质谱、薄层层析和生物降解鉴定琼胶寡糖的方法和依据,为后续研究琼胶酶的分类提供了技术支持与理论依据。 展开更多
关键词 琼寡糖 新琼寡糖 核磁 质谱 薄层层析
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Enhanced a novel β-agarase production in recombinant Escherichia coli BL21(DE3) through induction mode optimization and glycerol feeding strategy
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作者 CHAN Zhuhua CHEN Xinglin +4 位作者 HOU Yanping GAO Boliang ZHAO Chungui YANG Suping ZENG Runying 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2018年第2期110-118,共9页
Agarases are hydrolytic enzymes that act on the hydrolysis of agar and have a broad range of applications in food,cosmetics and pharmaceutical industries. In this study, a glycerol feeding strategy based on induction ... Agarases are hydrolytic enzymes that act on the hydrolysis of agar and have a broad range of applications in food,cosmetics and pharmaceutical industries. In this study, a glycerol feeding strategy based on induction mode optimization for high cell density and β-agarase production was established, which could effectively control acetate yield. First, exponential feeding strategy of glycerol with different overall specific growth rates(μ) was applied in the pre-induction phase. The results showed that the low μ(μ=0.2) was suggested to be the optimal for cell growth and β-agarase production. Second, the effects of induction temperature and the inducer concentration on cell growth and β-agarase production were investigated in the post-induction phase. When induced by isopropyl-β-d-thiogalactoside(IPTG), the strategy of 0.8 mmol/L IPTG induction at 20℃ was found to be optimal for β-agarase production. When cultivation was induced by continuous lactose feeding strategy of 1.0 g/(L·h), the β-agarase activity reached 112.5 U/mL, which represented the highest β-agarase production to date.Furthermore, the β-agarase was capable of degrading G. lemaneiformis powder directly to produce neoagarooligosaccharide, and the hydrolysates were neoagarotetraose(NA4) and neoagarohexaose(NA6). The overall research may be useful for the industrial production and application of β-agarase. 展开更多
关键词 β-agarase Escherichia coli process optimization glycerol feeding strategy neoagarooligosaccharide
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海洋紫色杆菌β-琼胶酶的分离纯化及性质 被引量:8
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作者 时岩玲 于文功 路新枝 《武汉大学学报(理学版)》 CAS CSCD 北大核心 2008年第4期497-502,共6页
从青岛近海海水中分离得到一株高产琼胶酶的海洋紫色杆菌SY12.紫色杆菌SY12的发酵液上清经硫酸铵沉淀,凝胶过滤层析,离子交换层析和凝胶过滤层析等蛋白纯化步骤,得到纯化的野生琼胶酶AgaY.琼胶酶纯化了133.63倍,酶比活力为320.7 U/mg;... 从青岛近海海水中分离得到一株高产琼胶酶的海洋紫色杆菌SY12.紫色杆菌SY12的发酵液上清经硫酸铵沉淀,凝胶过滤层析,离子交换层析和凝胶过滤层析等蛋白纯化步骤,得到纯化的野生琼胶酶AgaY.琼胶酶纯化了133.63倍,酶比活力为320.7 U/mg;纯化的琼胶酶经SDS-PAGE检测,显示为单一条带,其相对分子质量约为50×103.研究结果表明琼胶酶AgaY降解琼脂糖的β-1,4-糖苷键,其主要产物为新琼四糖和新琼二糖.对AgaY进行酶学性质研究,发现其最适反应pH为7.0,最适反应温度为40℃;Hg2+,Ag+,Zn2+,Cu2+和Pb2+等金属离子可以显著的抑制琼胶酶AgaY的活力.研究发现,AgaY的催化活性不依赖于离子的存在,AgaY对EDTA不敏感. 展开更多
关键词 紫色杆菌 Β-琼胶酶 分离纯化 新琼寡糖
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