costaricstreptomyces 45I-3在不同底物下深层发酵产木聚糖酶的优化

S. Sipriyadi, A. Wahyudi, M. Suhartono, A. Meryandini
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引用次数: 4

摘要

木聚糖酶是一种重要的水解酶,在许多工业中都有广泛的应用,但获得木聚糖酶衍生产品并不容易。因此,优化木聚糖酶的高效生产是生物技术应用的一个重要方向。本研究旨在确定S. costaricanus 45I-3产木聚糖酶的底物类型、培养基组成和最佳条件。通过在桦木木聚糖、山毛榉木聚糖、燕麦木聚糖、玉米芯木聚糖和烟草木聚糖底物上培养被试菌,确定底物类型,同时通过测定不同底物浓度下木聚糖酶活性,替换碳、氮、磷酸盐和表面活性剂源,确定培养基组成和产酶量。结果表明,玉米芯木聚糖底物的酶促指数最高,为3.60,山毛榉木聚糖底物的酶促指数次之,为2.87 EI,但山毛榉木聚糖底物纯度更高,因此选择该底物作为木聚糖源进行进一步优化测定。28ºC, 120转/分,接种后第8天木聚糖酶活性最高,为2.29 U/mL。阿拉伯糖作为碳源产生的活性最高,为3.161 U/mL,而磷酸的最优来源是Na2HPO4 (2.37 U/mL)。两种氮源即硫酸铵氮(NH4)2SO4和酵母提取物均能产生2.57和2.36 U/mL的木聚糖酶。在生产培养基中添加表面活性剂,SDS (0.146 U/mL)和Tween 80 (0.438 U/mL)表现出负响应,降低了活性。综上所述,在不同的C、N、P源条件下,优化后的木聚糖酶活性均有所提高,且利用氮源(NH4)2SO4代替氮源酵母抽提物更经济,可以降低木聚糖酶的生产成本。
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Optimization of Xylanase Production by Streptomyces costaricanus 45I-3 Using Various Substrates through Submerged Fermentation
Xylanase is an important hydrolytic enzymes with many application in several industries, but to obtain enzyme derived products is not easy. Thus, the optimization of efficient xylanases production is a great interest for biotechnological application. This study aims to determine the type of substrate, medium composition, and optimum conditions of xylanase production by S. costaricanus 45I-3. Determination of substrate type was done by growing the tested bacteria on birchwood xylan, beechwood xylan, oat spelled xylan, corn cobs xylan, and tobacco xylan substrate, meanwhile the determination of medium composition and enzyme production were done by measuring xylanase activity at various substrate concentration and replacing the carbon, nitrogen, phosphate and surfactants source. The results showed that the highest enzymatic index (EI) produced from corn cob xylan substrate at 3.60 meanwhile the second highest was beechwood xylan substrate at 2.87 EI, however this substrate is purer, thus this substrate was selected and used as xylan sources for further optimization measurement. The best xylanase activity (2.29 U/mL) obtained on eighth day after inoculation on rotary incubator at 120 rpm in 28 ºC. Arabinose as the source of carbon generate the highest activity at 3.161 U/mL meanwhile the most preferred source of phosphate is Na2HPO4 (2.37 U/mL). Both source of nitrogen i.e. nitrogen ammonium sulphate (NH4)2SO4 and yeast extract were able to produce xylanase at 2.57 and 2.36 U/mL. The addition of surfactant in production medium showed addition of SDS surfactant (0.146 U/mL) and Tween 80 (0.438 U/mL) showed a negative response by decreasing the activity. The conclusion showed that the xylanase activity was increased after optimization at various C, N, and P sources, and the use of nitrogen source (NH4)2SO4), become a more economical alternative to replacing a nitrogen source yeast extract so it can lower the production costs of xylanase enzyme.
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