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海洋真菌来源的几丁质酶生产 N-乙酰-D-氨基葡萄糖:利用废弃几丁质进行增值的潜在工业意义案例研究。

N-Acetyl-D-glucosamine Production by a Chitinase of Marine Fungal Origin: a Case Study of Potential Industrial Significance for Valorization of Waste Chitins.

机构信息

Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, West Bengal, 721302, India.

Advanced Laboratory for Plant Genetic Engineering, Advanced Technology Development Center, Indian Institute of Technology Kharagpur, Kharagpur, 721302, India.

出版信息

Appl Biochem Biotechnol. 2019 Jan;187(1):407-423. doi: 10.1007/s12010-018-2822-3. Epub 2018 Jul 2.

Abstract

Chitin is a linear homo-polymer of N-acetyl-D-glucosamine (GlcNAc) and the second most abundant biopolymer after cellulose. Several industries rely on the bioprocesses for waste chitin recycle and hydrolysis by chitinase (EC 3.2.1.14) for potential healthcare applications through the production of its monomeric subunit, GlcNAc. In the present study, a chitinase-producing fungus (named as MFSRK-S42) was isolated from the marine water sample of North Bay of the Andaman and Nicobar Islands. It was identified as Aspergillus terreus by morphological and molecular characterization methods leveraging the internal transcribed spacer between 18S rRNA and 5.8S rRNA. Chitinase that was isolated from the fermentation broth of marine Aspergillus terreus was used to carry out biotransformation of chitineaceous wastes. Prior to the enzymatic hydrolysis step, chitins from different sources were characterized for the presence of characteristic functional groups, grain size distribution, and surface morphology. Enzymatic hydrolysis of 50 mg/ml substrate with six units of enzyme incubated for 5 days revealed 15, 36.5, 40, and 46 mg/ml GlcNAc production from ground prawn shell, chitin flakes, colloidal prawn shell, and swollen chitin respectively under standardized conditions, as determined by HPLC. In this study, 30, 73, 80, and 92% GlcNAc yields were observed from ground prawn shell, chitin flakes, colloidal prawn shell, and swollen chitin conversion respectively. The HPLC-eluted product was confirmed as GlcNAc by the presence of characteristic functional groups in FTIR and 244 Da molecular weight peak in HRMS analyses.

摘要

甲壳素是一种由 N-乙酰-D-葡萄糖胺(GlcNAc)组成的线性均聚物,是仅次于纤维素的第二大丰富生物聚合物。有几个行业依赖于生物过程来回收废甲壳素,并通过甲壳素酶(EC 3.2.1.14)水解,以生产其单体亚基 GlcNAc,从而实现潜在的医疗保健应用。在本研究中,从安达曼和尼科巴群岛北湾的海水样本中分离出一种产甲壳素酶的真菌(命名为 MFSRK-S42)。通过形态学和分子特征鉴定方法,利用 18S rRNA 和 5.8S rRNA 之间的内部转录间隔区,将其鉴定为土曲霉。从海洋土曲霉发酵液中分离出的甲壳素酶用于进行甲壳质废物的生物转化。在进行酶水解步骤之前,对来自不同来源的甲壳素进行了特征官能团、粒度分布和表面形态的特性分析。在标准条件下,用 50mg/ml 底物和 6 个单位的酶孵育 5 天,对虾壳粉、甲壳素薄片、胶体虾壳和膨胀甲壳素进行酶解,分别得到 15、36.5、40 和 46mg/ml 的 GlcNAc。在这项研究中,分别从虾壳粉、甲壳素薄片、胶体虾壳和膨胀甲壳素的转化中观察到 30%、73%、80%和 92%的 GlcNAc 收率。通过 HPLC 洗脱产物在 FTIR 中存在特征官能团和 HRMS 分析中 244Da 分子量峰,证实其为 GlcNAc。

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