Table 1 Proteins whose total and/or functional yields increase upon co-production of GroESL

Human procollagenase

GroESL increased production levels by 10-fold, solubility and half-life

p50^csk Protein-Tyrosine Kinase

Co-production enhanced solubility and activity of the protein by up to >50%

Candida albicans PMI metalloenzyme

2-fold increase in protein solubility

α and β subunits of human propionyl-CoA carboxylase (PCC)

Several hundred-fold increase in PCC specific activity; most of the protein produced in soluble form

Human electron transfer flavoprotein (ETF)

Co-production required for stable expression of ETF α G116R mutant

β-glucosidase from Cellovibrio gilvus and Agrobacterium tumefaciens

Co-production resulted in slower growth rate and reduced yield but increased solubility of the proteins by 20–60% at 37°C and up to 70% at 25°C

Cryj2 Japanese cedar pollen

Increased yield and solubility of expressed protein; 4-fold stabilisation of the protein in the presence of a 10-fold chaperone excess

Human kinase inhibitor-GST fusion

Solubility of otherwise mostly insoluble protein enhanced by 5–6-fold

Eukaryotic phenylalanine ammonia-lyase

Dramatically improved yield and activity of the protein after engineering of gene to remove E. coli rare codons

Bovine adrenodoxin reductase (AdR)

Increased soluble AdR yield to 10 mg/l, compared with 4 mg/l with Hsp70

Cyanobacterium transcription factor

3–4-fold increase in solubility

Thermococcus litoralis 4-α-glucanotransferase (GTase)

Co-production of GroESL with tRNA_AGA and tRNA_AGG led to 5-fold increase in GTase activity in soluble fraction; yield otherwise lower and 60% insoluble

Agrobacterium radiobacter carbamoylase

4-fold increase in activity due to improved solubility

Human cytochrome P450 3A7 (CYP3A7)

Increased expression levels and activity of the otherwise inactive protein

Decarboxylase component of human α-keto acid dehydrogenase complex

Co-production of GroEL or GroES resulted in increase in decarboxylase activity by 500-fold and 30-fold, respectively

Maize plastidic protoporphyrinogen IX oxidase (PPO)

6-fold increase in soluble PPO yield

Manganese catalase from Thermus sp.

Increased solubility (up to 50%) with GroESL

p66 and p51 subunits of HIV-1 RTase

Yield and nucleic acid affinity increased by 4–5- and 1.6-fold, respectively

Anti-digoxin Fab antibody fragment

4-fold increase in solubility of the Fab produced in E. coli Origami strain

Agrobacterium tumefaciens D-carbamoylase (DCB)

Increase in solubility of DCB up to 60% and activity by 6.2-fold at 28°C; at 25°C protein solubility increased to 75% and activity by 4.5-fold

Guinea pig NADPH:quinone oxidoreductase

3-fold increase in solubility

Aconitase

Solubility and activity increased to 40% and by 1.5-fold, respectively

Rhodococcus erythropolis desulfinase

Solubility of the protein increased up to 40–50% and activity by 25-fold

E. coli glutamate racemase (GluR)

Growth of host cells improved; 2.2-fold increase in yield of active GluR

Pseudomonas putida F61 nicotinoprotein formaldehyde dismutase (NDF)

With tac promoter, increased solubility (up to 80%) and 6-fold higher enzyme activity; lesser effect when NDF expressed under the lac promoter

Human PP2A methyltransferase

24-fold increase in solubility

Oligo-1,6-glucosidase from Bacillus thermoglucosidasius

Specific activity increased by 44%, 56% and 56% with co-production of GroES, GroEL and GroESL, respectively

Cyclodextrin gluanotransferase (CGTase) from Bacillus macerans

Increase in solubility and activity of CGTase by 12% and 1.5-fold, respectively, at 37°C and by 22% and 1.3-fold, respectively, at 25°C

Rhizobium sp. α-1,6-fucosyltransferase

At 30°C, improved folding and an increase in specific activity by 1.76-fold

Mouse CYP27B1 protein

10-fold increase in the yield of stable and active protein

Pyridoxine 4-oxidase (PNO) from Microbacterium luteolum

No benefit of GroESL at 37°C; co-production at 23°C enhanced solubility and specific activity of PNO by 1.9-fold and 3.9-fold, respectively

Pyridoxal 4-dehydrogenase from Microbacterium luteolum

Co-production at 20°C led to reduced amounts of insoluble protein and increased specific activity by 9.1-fold

Alcaligenes xylosoxydans N-acyl-D-amino acid amidohydrolases

Enzymatic activity of the proteins increased from 7.8 to 72.4 U/mg and 7.1 to 22.7 U/mg, respectively, at 30°C

Human aromatase (P450arom, CYP19) NmA264C and NmA264R mutants

No improvement with NmA264C; production of NmA264R greatly enhanced (up to 400 nmol/l)

scFv specific for c-Met

Solubility increased 2-fold in E. coli Origami2(DE3) but not in BL21(DE3)

Yeast mitochondrial aconitase

Increased solubility at 25°C with no change in total yield

human prolyl hydroxylase isoenzyme

2-fold increase in solubility when produced at 30°C

Pig liver esterase γ-isoenzyme (PLE)

Enhanced yield of soluble and active PLE in E. coli Origami (DE3)

Soybean seed ferritin complex

Increased solubility of H-1 subunit from 4 to 39% and H-2 subunit from 19 to 85%

Human 11β hydroxylase

20- to 40-fold increase in yield in half the production time

Human glucose 6-phosphate dehydrogenase (G6PD) and mutants

Negligible effect on expression of wild type G6PD but activities of two mutants were enhanced by 48–160% and 39–118% at 37°C and 31°C, respectively