Poonsrisawat Aphisit, Paemanee Atchara, Wanlapatit Sittichoke, Piyachomkwan Kuakoon, Eurwilaichitr Lily, Champreda Verawat
Enzyme Technology Laboratory, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Klong Luang, 12120 Pathum Thani Thailand.
Genome Institute, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Klong Luang, 12120 Pathum Thani Thailand.
3 Biotech. 2017 Oct;7(5):290. doi: 10.1007/s13205-017-0924-1. Epub 2017 Aug 28.
In this study, an efficient ethanol production process using simultaneous saccharification and viscosity reduction of raw cassava pulp with no prior high temperature pre-gelatinization/liquefaction step was developed using a crude starch- and cell wall-degrading enzyme preparation from BCC17849. Proteomic analysis revealed that the enzyme comprised a complex mixture of endo- and exo-acting amylases, cellulases, xylanases, and pectina ses belonging to various glycosyl hydrolase families. Enzymatic hydrolysis efficiency was dependent on the initial solid loading in the reaction. Reduction in mixture viscosity was observed with a rapid decrease in complex viscosity from 3785 to 0.45 Pa s with the enzyme dosage of 2.19 mg/g on a dried weight basis within the first 2 h, which resulted from partial destruction of the plant cell wall fiber and degradation of the released starch granules by the enzymes as shown by scanning electron microscopy. Saccharification of cassava pulp at an initial solid of 16% (w/v) in a bench-scale bioreactor resulted in 736.4 mg glucose/g, which is equivalent to 82.92% glucose yield based on the total starch and glucan in the substrate, after 96 h at 40 °C. Simultaneous saccharification and fermentation of cassava pulp by with the uncooked enzymatic process led to a final ethanol concentration of 6.98% w/v, equivalent to 96.7% theoretical yield based on the total starch and cellulose content. The results demonstrated potential of the enzyme for low-energy processing of cassava pulp in biofuel industry.
在本研究中,利用来自BCC17849的粗淀粉和细胞壁降解酶制剂,开发了一种高效的乙醇生产工艺,该工艺对木薯原浆进行同步糖化和降低粘度,无需事先进行高温预糊化/液化步骤。蛋白质组学分析表明,该酶由属于不同糖基水解酶家族的内切和外切淀粉酶、纤维素酶、木聚糖酶和果胶酶的复杂混合物组成。酶促水解效率取决于反应中的初始固体负载量。在最初2小时内,随着酶用量为2.19mg/g(基于干重),混合液粘度迅速从3785降至0.45Pa·s,观察到混合液粘度降低,这是由于植物细胞壁纤维的部分破坏以及扫描电子显微镜所示的酶对释放出的淀粉颗粒的降解所致。在实验室规模的生物反应器中,以16%(w/v)的初始固体含量对木薯原浆进行糖化,在40℃下96小时后,得到736.4mg葡萄糖/g,相当于基于底物中总淀粉和葡聚糖的葡萄糖产率为82.92%。采用未蒸煮酶法对木薯原浆进行同步糖化和发酵,最终乙醇浓度达到6.98%(w/v),相当于基于总淀粉和纤维素含量的理论产率的96.7%。结果证明了该酶在生物燃料工业中对木薯原浆进行低能耗加工的潜力。