Table 3.
Examples for one- to two step enzymatic reactions employing engineered yeast whole-cell biocatalysts (+ overexpression; - knockout/deletion).
| Whole-Cell Biocatalyst |
Engineering + overexpression - deletion |
Substrate | Product | Performance: yield (ee) | Ref. | ||
| Saccharomyces cerevisiae | - β-keto reductase |  |
 |
53% (90%) (slow, continuous substrate feed) | [151] | ||
| Saccharomyces cerevisiae | A: +Fasp, -Gre2p, -Ypr1pIV |  |
 |
A: | d: 78% (91% R)II | [152] | |
| B: -Fasp, +Gre2pIV | e: 26% (> 98% RI)I | ||||||
| a: R1 = Me; R3 = Me | B: | a: 76% (> 98% S)I | |||||
| b: R1 = Et; R3 = Me | b: 85% (> 98% S)I | ||||||
| c: R1 = Me; R3 = Et | c: 83% (> 98% S)I | ||||||
| d: R1 = Et; R3 = Et | d: 87% (> 98% S)I | ||||||
| e: R1 = n-Pr; R3 = Et | e: 90% (> 98% S)I | ||||||
| Saccharomyces cerevisiae | A: - Gre2p, + Ypr1pIV |  |
a: R2 = Me |  |
A: | a: 90% (98% syn)II | [152] |
| B: + Gre2P, - Ypr1pIV | b: R2 = Et | b: 89% (83% syn)II | |||||
| c: R2 = allyl | c: 92% (65% syn)II | ||||||
| d: R2 = propargyl | d: 75% (> 98% syn)II | ||||||
| B: | a: 86% (70% syn)I | ||||||
| b: 73% (67% anti)I | |||||||
| c: 67% (> 98% anti)I | |||||||
| d: 70% (> 98% anti)I | |||||||
| Saccharomyces cerevisiae | A: +Ymr226cIII |  |
 |
A: 72% (87%, S)I | [153,154] | ||
| B: +Ara1pIII | R1 = Me; R2 = Me | B: 48% (91%, S)I | |||||
| C: +Ypr1pIII | C: 62% (87%, S)I | ||||||
| Saccharomyces cerevisiae | + reductase gene YDR368wV |
 bicyclo[2.2.2]octane-2,6-dione |
 (1R,4S,6S)-6-hydroxy-bicyclo[2.2.2]octane-2-ol |
97% de | [155] | ||
| > 99% ee | |||||||
| 84% yield | |||||||
| Saccharomyces cerevisiae | + multiple copies of Candida tenuis xylose reductase variant W23F |  |
a: R = H |  |
a: 35% (> 99.9%, S)VI | [158] | |
| b: R = Ph | b: 80% (> 99.9%, R)VI | ||||||
| c: R = o-Cl-Ph | c: 76% (99.6%, R)VI | ||||||
| d: R = m-Cl-Ph | d: 14% (> 99.9%, R)VI | ||||||
| e: R = p-Cl-Ph | e: 74% (99.9%, R)VI | ||||||
| f: R = p-CN-Ph | f: 100% (99.4%, n.d.)VI | ||||||
| Saccharomyces cerevisiae | + Antirrhinum majus benzoic acid methyltransferase (BAMT) |  |
 |
~1 mg methyl benzoate per L of culture (with OD600 = 1), after 24 h | [160] | ||
| Pichia pastoris | + Rhodotorula glutinis epoxide hydrolase |  |
 |
~36% (> 98%) (theoretical yield: 50%) | [171] | ||
| Schizosaccharomyces pombe | + human CYP11B1-V78I + electron transfer proteins adrenodoxin (Adx) & adrenodoxin reductase (AdR) |
 |
 |
2.5-fold higher activity compared with parental strain | [172] | ||
| Schizosaccharomyces pombe | + human CYP2D6 |  |
 |
56% yield 98.5% purity |
[173] | ||
| Schizosaccharomyces pombe | + Trigonopsis variabilis D-Amino Acid Oxidase (multi copies) - catalase |
Cephalosporin C | α-ketoadipyl-7-cephalosporanic acid and glutaryl-7-amino-cephalosporanic acid | 550 U/g CDW increased mechanical resistance | [191] | ||
Icarbon source: glucose
IIcarbon source: galactose
IIIAra1p = S. cerevisiae reductase, NADPH-dependent; Ypr1p = S. cerevisiae reductase, NADPH-dependent; YMR226c = S. cerevisiae short chain dehydrogenase ORF [153].
IVFasp = S. cerevisiae fatty acid synthase; Gre2p = S. cerevisiae α-acetoxy ketone reductase; Ypr1p = S. cerevisiae aldo-keto reductase [152].
VS. cerevisiae 2-methylbutyraldehyde reductase (NCBI accession number: NP 010656)
VIWhole-cell bioreductions of α-keto esters (10 mM) under anaerobic conditions using 1 M ethanol as co-substrate; n.d. = not determined.