哈茨木霉生产β-(1,3)-葡聚糖酶的研究:以帕拉蒙和普斯图兰为原料生产低聚糖的优化与应用

E. Giese, R. Dekker, A. M. Barbosa, M. L. C. Silva, R. Silva
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引用次数: 16

摘要

以哈茨木霉(Trichoderma harzianum Rifai) PAMB-86为原料,利用响应面法对β-(1→3)-葡聚糖酶进行了优化。酶滴度最大值出现在第5天,初始pH为5.5,曝气1.5vvm。通过凝胶过滤将T. harzianum Rifai PAMB- 86产生的β-(1→3)- β-葡聚糖水解酶复合物分离成2个部分(F-I, F-II),分别从海藻paramylon((1→3)-β- d -葡聚糖)和地衣pustulan((1→6)-β- d -葡聚糖)中制备低聚糖。这两种酶在30 min内对paramylon的攻击程度为~15-20%,最终产物为葡萄糖和层状糖,以及聚合度(DP)≥3的层状糖-寡糖。只有f - 1在30 min内降解了pustulan,降解率约为2%,主要产物为葡萄糖、龙胆糖和DP≥4的龙胆寡糖。水解产物性质的差异可以用每个酶组分的底物特异性和攻击的β- d -葡聚糖的结构差异来解释。
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Production of β-(1,3)-glucanases by Trichoderma harzianum Rifai: Optimization and Application to Produce Gluco-oligosaccharides from Paramylon and Pustulan
β-(1→3)-Glucanases were produced by Trichoderma harzianum Rifai PAMB-86 cultivated on botryosphaeran in a bench-fermenter and optimised by the response surface method. Maximal enzyme titres occurred at 5 days, initial pH 5.5 and aeration of 1.5vvm. β-(1→3)-The β-glucanolytic enzyme complex produced by T. harzianum Rifai PAMB- 86 was fractionated by gel filtration into 2 fractions (F-I, F-II), and employed to produce gluco-oligosaccharides from algal paramylon ((1→3)-β-D-glucan) and lichen pustulan ((1→6)-β-D-glucan). Both enzymes attacked paramylon to the extent of ~15-20% in 30 min releasing glucose and laminaribiose as major end-products, and laminari- oligosaccharides of degree of polymerization (DP) ≥ 3. Only F-I degraded pustulan resulting in ~2% degradation at 30 min, with glucose, gentiobiose and gentio-oligosaccharides of DP ≥ 4 as major products. The difference in the nature of the hydrolysis products can be explained by the substrate specificities of each enzyme fraction, and the structural differences of the β-D-glucans attacked.
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