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

BMC Energy

Table 2 Recent studies of protein engineering in biofuel research

Enzyme type	Target property	Screening assay
Biomass deconstruction
Endoglucanase	Activity	Chromogenic surrogate substrate [40]
Endoglucanase	Thermostability	Colorimetric assay on polymerization degree [41]
Cellobiohydrolase	Activity/thermostability	Colorimetric assay on reducing sugar [42]
Beta-glucosidase	Activity	Chromogenic surrogate substrate [43]
Beta-glucosidase	Activity	MALDI MS imaging [44]
Endoxylanase	Activity	Colorimetric assay on reducing sugar [45]
Endoxylanase	Product inhibition	Chromogenic surrogate substrate [46]
Endoxylanase	Thermostability	Colorimetric assay on reducing sugar [47]
Substrate utilization
Glucose oxidase	Stability	Fluorescent redox indicator [48]
Cellobiose dehydrogenase	Activity	Fluorescent redox indicator [49]
Xylose reductase	Cofactor specificity	Colorimetric assay on NAD(P)H consumption [50, 51]
Xylose transporter	Activity	Genetic biosensor [9]
Product synthesis
P450 fatty acid decarboxylases	Substrate specificity	GC-MS [52]
Citramalate synthase	Activity for C3/C4 alcohol production	Synthetic auxotrophy [53]
2-Isopropylmalate synthase Ketoisovalerate decarboxylase	Substrate specificity towards C5–C8 alcohol production	GC-MS [54]
Fatty acid synthase	Product specificity towards C6/C8 fatty acids	GC-MS [55]
Thioesterase	Product specificity towards C8/C12 fatty acids	GC-MS [56]
Thioesterase	Product specificity towards C8 fatty acid	Synthetic auxotrophy [57]
Lipase	Activity/substrate specificity/stability	Chromogenic surrogate substrate [58,59,60]
Lipase	Enantioselectivity	GC [61]

ISSN: 2524-4469