Table 4.

Recombinant Protein production using methylotrophs.

Protein	Strain	Methanol assimilation pathway	Metabolic engineering strategy	Substrate	Culture condition	Product titer	Ref.
Methylotrophic Yeasts
Hepatitis B surface antigen (HBsAg)	P. pastoris	XuMP	Expression of HBsAg under AOX1 promoter	Methanol and glycerol	Chemostat fermentation using glycerol to obtain high cell density followed by methanol induction	187.71 mg/l	[69]
Trypsinogen	P. pastoris	XuMP	Expression of porcine trypsinogen using AOX1 promoter	Methanol and glucose	Continuous chemostat fermentation using glucose and methanol	210 mg/l	[67]
Cellulase	P. pastoris	XuMP	TrCBH2 encoding cellulase under DES promoter	Methanol and glycerol	Two-stage Fed-batch fermentation using glycerol to obtain high cell density followed by methanol induction	18 g/l	[140]
Human serum albumin (HSA)	P. pastoris	XuMP	Expression of Hsa gene under AOX1 promoter	Methanol and glycerol	Two-stage fed-batch fermentation using glycerol for growth followed by methanol induction for 96 h	8.86 g/l	[141]
	P. pastoris (ALE-based methanol tolerant strain)	XuMP	Expression of Has gene under AOX1 promoter in Adapted strain	Methanol and glucose	Two-stage fed-batch fermentation using glucose for growth followed by methanol induction for 48 h	350 mg/l	[142]
Plectasin	P. pastoris	XuMP	Expression of gene encoding plectasin	Methanol and glycerol	Two-stage fed-batch fermentation using glycerol for growth followed by methanol induction for 120 h	748.6 μg/ml	[143]
Xylanase	P. pastoris	XuMP	Heterologous expression of xyn11A under AOX1 promoter	Methanol	Fed-batch using methanol as the sole carbon source	1.16 g/l	[71]
Methylotrophic bacteria
Green fluorescent protein (GFP)	M. extorquens AM1	Serine cycle	Heterologous expression of GFP under mxaF promoter	Methanol	Fed-batch using methanol as sole carbon source	4 g/L	[72]
Enterocin P	M. extorquens AM1	Serine cycle	Heterologous expression of EntP under mxaF promoter	Methanol	Shake flask using CHOI-medium containing methanol as sole carbon source	155 ng/ml	[73]