rapid optimization of enzyme mixtures for deconstruction of diverse pretreatmentbiomass feedstock combinations解构的快速优化酶混合物不同pretreatmentbiomass原料组合.pdfVIP

Banerjee et al. Biotechnology for Biofuels 2010, 3:22 /content/3/1/22 RESEARCH Open Access Rapid optimization of enzyme mixtures for deconstruction of diverse pretreatment/biomass feedstock combinations * Goutami Banerjee, Suzana Car, John S Scott-Craig, Melissa S Borrusch, Jonathan D Walton Abstract Background: Enzymes for plant cell wall deconstruction are a major cost in the production of ethanol from lignocellulosic biomass. The goal of this research was to develop optimized synthetic mixtures of enzymes for multiple pretreatment/substrate combinations using our high-throughput biomass digestion platform, GENPLAT, which combines robotic liquid handling, statistical experimental design and automated Glc and Xyl assays. Proportions of six core fungal enzymes (CBH1, CBH2, EG1, b-glucosidase, a GH10 endo-b 1,4-xylanase, and b- xylosidase) were optimized at a fixed enzyme loading of 15 mg/g glucan for release of Glc and Xyl from all combinations of five biomass feedstocks (corn stover, switchgrass, Miscanthus, dried distillers’ grains plus solubles [DDGS] and poplar) subjected to three alkaline pretreatments (AFEX, dilute base [0.25% NaOH] and alkaline peroxide [AP]). A 16-component mixture comprising the core set plus 10 accessory enzymes was optimized for three pretreatment/substrate combinations. Results were compared to the performance of two commercial enzymes (Accellerase 1000 and Spezyme CP) at the same protein loadings. Results: When analyzed with GENPLAT, corn stover gave the highest yields of Glc with commercial enzymes and with the core set with all pretreatments, whereas corn stover, switchgrass and Miscanthus gave comparable Xyl yields. With commercial enzymes and with the core set, yi