一种用于估计微生物降解几丁质废物产生的壳聚糖的简单快速比色法。
A simple and rapid colorimetric method for the estimation of chitosan produced by microbial degradation of chitin waste.
机构信息
School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha 751024, India.
School of Biotechnology, KIIT Deemed to be University, Bhubaneswar, Odisha 751024, India; School of Law, KIIT Deemed to be University, Bhubaneswar, Odisha 751024, India.
出版信息
J Microbiol Methods. 2019 Mar;158:66-70. doi: 10.1016/j.mimet.2019.02.001. Epub 2019 Feb 3.
Chitin is one of the most abundant biopolymers present in the environment. Chitosan being its major derivative can be obtained by hydrolysis of chitin, especially by microbial degradation. Estimation of resulting chitosan produced by chitin degradation is crucial to the process. Usefulness of the method of Badawy (Badawy, 2012) for estimation of chitosan is limited by interference resulting from susceptibility to variation in the pH of the sample and thiobarbituric acid. This work presents an improvement of the method proposed by Badawy for colorimetric determination of chitosan by using 3, 5-Dinitrosalicylic acid (DNSA) reagent instead of thiobarbituric acid, after one step depolymerization and deamination of chitosan with sodium nitrite (NaNO). Eventually colorimetric estimation was carried out at 540 nm. With the use of DNSA reagent, the limitation of thiobarbituric acid are overcome. This method is easy, cost effective, and sensitive for quantitative determination of chitosan. This new improved method was applied for evaluation and quantification of chitosan produced by microbial degradation of chitin waste by different novel Streptomyces strains.
几丁质是存在于环境中的最丰富的生物聚合物之一。壳聚糖是其主要衍生物，可以通过几丁质的水解获得，特别是通过微生物降解。估计几丁质降解产生的壳聚糖对于该过程至关重要。Badawy 方法（Badawy，2012）用于估计壳聚糖的用途受到样品 pH 值变化和硫代巴比妥酸易受干扰的限制。本工作提出了一种改进的方法，通过使用 3,5-二硝基水杨酸（DNSA）试剂代替硫代巴比妥酸，在一步用亚硝酸钠（NaNO）进行壳聚糖的解聚和脱氨后，对壳聚糖进行比色测定。最终在 540nm 处进行比色估计。使用 DNSA 试剂可以克服硫代巴比妥酸的限制。该方法简单、经济实惠，对壳聚糖的定量测定具有较高的灵敏度。该新改进的方法已应用于不同新型链霉菌菌株对几丁质废物进行微生物降解产生的壳聚糖的评估和定量。
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