从内生链霉菌(Streptomyces mobaraensis DB13)中纯化和表征与木薯(Costus speciosus)相关的中等恒温生淀粉消化α-淀粉酶。

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General and Applied Microbiology Pub Date : 2024-05-02 Epub Date: 2023-08-25 DOI:10.2323/jgam.2023.08.001
Dina Barman, Mamtaj S Dkhar
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引用次数: 0

摘要

众所周知,内生放线菌能产生各种具有潜在工业应用价值的酶。α-淀粉酶是一类重要的工业酶,具有多方面的用途。本实验旨在表征一种产生中度恒温α-淀粉酶的内生链霉菌(Streptomyces mobaraensis DB13)的特性,该链霉菌分离自Costus speciosus (J. Koenig) Sm.该酶通过 60% 硫酸铵沉淀、透析和 Sephadex G-100 柱层析进行纯化。根据 12% SDS-PAGE,纯化的 α 淀粉酶分子量估计为 55 kDa。α-淀粉酶的最大活性在 pH 7.0、50°C 条件下达到,在 pH 7.0 和 8.0 条件下培养 2 小时后,α-淀粉酶的活性仍保持 80%。在 Tween-80、SDS、PMSF 和 Triton X-100 的存在下,α-淀粉酶的活性保持稳定;但是,β-巯基乙醇、EDTA 和 H2O2 会降低α-淀粉酶的活性。经计算,该α-淀粉酶的动力学参数 Km 和 Vmax 值分别为 2.53 mM 和 29.42 U/mL。该α-淀粉酶在 pH 值为 7.0、温度为 50℃、浓度为 10 mg/mL 的条件下具有消化各种生淀粉的能力,其中玉米和大米是首选底物。培养 4 小时后开始消化,培养 48 小时后达到最大消化率。这些结果表明,S. mobaraensis DB13 是中度恒温α-淀粉酶的潜在来源,可有效水解生淀粉。这表明这种α-淀粉酶有望用于工业用途。
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Purification and characterization of moderately thermostable raw-starch digesting α-amylase from endophytic Streptomyces mobaraensis DB13 associated with Costus speciosus.

Endophytic actinobacteria are known to produce various enzymes with potential industrial applications. Alpha-amylase is an important class of industrial enzyme with a multi-dimensional utility. The present experiment was designed to characterize a moderately thermostable α-amylase producing endophytic Streptomyces mobaraensis DB13 isolated from Costus speciosus (J. Koenig) Sm. The enzyme was purified using 60% ammonium sulphate precipitation, dialysis, and Sephadex G-100 column chromatography. Based on 12% SDS-PAGE, the molecular weight of the purified α-amylase was estimated to be 55 kDa. The maximum α-amylase activity was achieved at pH 7.0, 50°C and it retained 80% of its activity at both pH 7.0 and 8.0 after incubation for 2 h. The α-mylase activity is strongly enhanced by Ca2+, Mg2+, and inhibited by Ba2+. The activity remains stable in the presence of Tween-80, SDS, PMSF, and Triton X-100; however, β-mercaptoethanol, EDTA, and H2O2 reduced the activity. The kinetic parameters Km and Vmax values for this α-amylase were calculated as 2.53 mM and 29.42 U/mL respectively. The α-amylase had the ability to digest various raw starches at a concentration of 10 mg/mL at pH 7.0, 50°C, where maize and rice are the preferred substrates. The digestion starts after 4 h of incubation, which reaches maximum after 48 h of incubation. These results suggest that S. mobaraensis DB13 is a potential source of moderately thermostable α-amylase enzyme, that effciently hydrolyzes raw starch. It suggesting that this α-amylase is a promising candidate to be use for industrial purposes.

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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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