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Table 2 Recent studies of protein engineering in biofuel research

From: Recent advances in high-throughput mass spectrometry that accelerates enzyme engineering for biofuel research

Enzyme typeTarget propertyScreening assay
Biomass deconstruction
 EndoglucanaseActivityChromogenic surrogate substrate [40]
 EndoglucanaseThermostabilityColorimetric assay on polymerization degree [41]
 CellobiohydrolaseActivity/thermostabilityColorimetric assay on reducing sugar [42]
 Beta-glucosidaseActivityChromogenic surrogate substrate [43]
 Beta-glucosidaseActivityMALDI MS imaging [44]
 EndoxylanaseActivityColorimetric assay on reducing sugar [45]
 EndoxylanaseProduct inhibitionChromogenic surrogate substrate [46]
 EndoxylanaseThermostabilityColorimetric assay on reducing sugar [47]
Substrate utilization
 Glucose oxidaseStabilityFluorescent redox indicator [48]
 Cellobiose dehydrogenaseActivityFluorescent redox indicator [49]
 Xylose reductaseCofactor specificityColorimetric assay on NAD(P)H consumption [50, 51]
 Xylose transporterActivityGenetic biosensor [9]
Product synthesis
 P450 fatty acid decarboxylasesSubstrate specificityGC-MS [52]
 Citramalate synthaseActivity for C3/C4 alcohol productionSynthetic auxotrophy [53]
 2-Isopropylmalate synthase
Ketoisovalerate decarboxylase
Substrate specificity towards C5–C8 alcohol productionGC-MS [54]
 Fatty acid synthaseProduct specificity towards C6/C8 fatty acidsGC-MS [55]
 ThioesteraseProduct specificity towards C8/C12 fatty acidsGC-MS [56]
 ThioesteraseProduct specificity towards C8 fatty acidSynthetic auxotrophy [57]
 LipaseActivity/substrate specificity/stabilityChromogenic surrogate substrate [58,59,60]
 LipaseEnantioselectivityGC [61]