[논문]Medium optimization for keratinase production by a local Streptomyces sp. NRC 13S under solid state fermentation

Shata, Hoda Mohamed Abdel Halim; Farid, Mohamed Abdel Fattah

doi:10.3839/jabc.2013.020

[국내논문] Medium optimization for keratinase production by a local Streptomyces sp. NRC 13S under solid state fermentation 원문보기

Journal of applied biological chemistry, v.56 no.2, 2013년, pp.119 - 129

Shata, Hoda Mohamed Abdel Halim (Microbial Chemistry Department, National Research Center) , Farid, Mohamed Abdel Fattah (Natural and Microbial Products Department, National Research Center)

Abstract ▼ AI-Helper

Thirteen different Streptomyces isolates were evaluated for their ability to produce keratinase using chicken feather as a sole carbon and nitrogen sources under solid state fermentation (SSF). Streptomyces sp. NRC 13S produced the highest keratinase activity [1,792 U/g fermented substrate (fs)]. The phenotypic characterization and analysis of 16S rDNA sequencing of the isolate were studied. Optimization of SSF medium for keratinase production by the local isolate, Streptomyces sp. NRC13S, was carried out using the one-variable-at-a-time and the statistical approaches. In the first optimization step, the effect of incubation period, initial moisture content, initial pH value of the fermentation medium, and supplementation of some agro-industrial by-products on keratinase production were evaluated. The strain produced about 2,310 U/gfs when it grew on chicken feather with moisture content of 75% (w/w), feather: fodder yeast ratio of 70:30 (w/w), and initial pH 7 using phosphate buffer after 8 days. Based on these results, the Box-Behnken design and response surface methodology were applied to find out the optimal conditions for the enzyme production. The corresponding maximal production of keratinase was about 2,569.38 U/gfs.

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제안 방법

This report specifies the details of screening, identification of the most potent isolate for keratinase activity by phenotypic and 16S rDNA, and investigating some factors affecting enzyme production using the one-variable-at-a-time. Subsequently, the factors that had significant effects were optimized using BoxBehnken design.
4, respectively. Supplementation the feather medium with different agro-industrial by-products, wheat bran (WB), corn cobs (CC) rice husk (RH), rice straw (RS), fodder yeast (FY), earn cotton seed (ECS) and earn flaxseed (EFS), were tested and the best substrate for production of keratinase was supplemented in different concentrations. For each experimental variable, all other parameters were kept at their optimal level.
(i=1, 3; j=1, 3, i j) represent the independent variables in the form of coded values. Statistical software package Design-Expert (Version 8.0.2, State-Ease, Minneapolis, MN, USA) was used to design and analyze the experiment. A 2³ factorial design, with five replicates at the centre point with total number of 17 trials, was employed.

대상 데이터

2, State-Ease, Minneapolis, MN, USA) was used to design and analyze the experiment. A 2³ factorial design, with five replicates at the centre point with total number of 17 trials, was employed. The accuracy and general ability of the above polynomial model could be evaluated by the coefficient of determination R².

이론/모형

Box-Behnken design. Based on the results of the one-variable-at-a-time experiment response surface methodology (RSM) was used to optimize keratinase production using Box-Behnken design (Box and Behnken, 1960). The behavior of the system was explained by the following quadratic equation,
Protein assay. The protein content of the enzyme preparation was estimated by Lowry method (Lowry et al., 1951).

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