Solid-State Fermentation of Agroindustrial Residues for Glucoamylase Production from Endophytic Fungi Penicillium javanicum of Solanum tuberosum L.

M. El-Gendy, N. H. Alzahrani
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Abstract

Glucoamylase production has been evaluated under solid-state fermentation of agro-industrial residues including groundnut shell, corncob, corn stover, sugarcane bagasse, wheat straw, barely straw and rice straw as renewable cheap substrates by different 14 endophytic fungal species. Among them the endophytic fungi Penicillium javanicum obtained from the root of Solanum tuberosum L. showed the maximum yield of glucoamylase using groundnut shell as solid substrate (289.23 ± 0.80 U/gds). Under the optimized production parameters in solid state fermentation process (250 mL Erlenmeyer flask containing 20 grams groundnut shell supplemented with 30% soya waste as an inexpensive, eco-friendly way of enzyme production sieved to 1mm, moistened to 55% initial moisture content with potato process wastewater, pH 5.0, inoculum intense 2 × 108 spore and incubated at 30°C for 5 days fermentation period), a fourfold increase (4.19-fold) in glucoamylase production was occurred. In our study there was a strong relation between the enzyme secretion and the trophophase. The purified enzyme exhibited specific activity 81.60 and 237. 24 U/mg with enzyme recovery equal to 51.11 and 22.14% and purification fold 2.2 and 6.39-fold after the precipitation with (NH4)2SO4 and gel fractionation on sephadex G-100, respectively with maximum activity at 40-50°C and pH 5 and it was stable and retained 100% of its activity at temperature up to 60°C along with pH 5–7. The enzyme was not metallo enzyme due to EDTA and EGTA at 50 mM had no effect on glucoamylase activity but it was considered as a serine protease due to it lost 68 and 92% of its activity the serine protease inhibitor paramethyl sulfonyl fluoride (PMSF) at 10 and 50 mM, respectively.
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龙葵内生真菌爪哇青霉固态发酵农用残留物生产葡萄糖淀粉酶的研究。
以花生壳、玉米芯、玉米秸秆、甘蔗渣、小麦秸秆、秸秆和水稻秸秆为可再生廉价底物,对14种不同内生真菌在固态发酵条件下的葡萄糖淀粉酶产量进行了研究。其中以落地壳为固体底物产糖淀粉酶的内生真菌javanicum产糖淀粉酶最高(289.23±0.80 U/gds)。在优化的固体发酵工艺条件下(250 mL Erlenmeyer烧瓶中含有20 g花生壳,添加30%大豆废弃物,筛选至1mm,用马铃薯加工废水浸湿至初始含水量55%,pH为5.0,接种强度为2 × 108个孢子,在30℃发酵5天),葡萄糖淀粉酶产量提高4倍(4.19倍)。在我们的研究中,酶的分泌与滋养期有很强的关系。纯化酶的比活性分别为81.60和237。在sephadex G-100上经(NH4)2SO4沉淀和凝胶分离,酶回收率分别为51.11和22.14%,纯化倍数分别为2.2和6.39倍,在40-50°C和pH 5条件下活性最高,在60°C和pH 5 - 7条件下稳定且保持100%的活性。由于EDTA和EGTA在50 mM时对葡萄糖淀粉酶活性没有影响,因此该酶不是金属酶,但由于其在10和50 mM时分别失去了丝氨酸蛋白酶抑制剂PMSF的68%和92%的活性,因此被认为是丝氨酸蛋白酶。
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