Table 1 Top-down genome minimisation projects

From: Trimming the genomic fat: minimising and re-functionalising genomes using synthetic biology

Parental strain and size

Designation and size

Minimisation method

Size/ Percentage of reduction

Phenotypes of the resulting strains

Applications

Refs

Escherichia coli W3110 4.65 Mb

E. coli MGF-01 3.62 Mb

Lambda homologous recombination and P1 transduction

1030 kb (22.2%)

Improved growth in M9 minimal medium

Enhanced threonine production

11,15

Escherichia coli W3110 4.65 Mb

E. coli DGF-298 2.98 Mb

Lambda homologous recombination and P1 transduction

1670 kb (35.9%)

Better growth fitness and cell yield in a rich medium

N/A

16

Escherichia coli MG1655 4.64 Mb

E. coli CDΔ3456 4.33 Mb

Tn5-targeted Cre/loxP excision and P1 transduction

313.1 kb (6.7%)

Normal growth in LB broth

N/A

9

Escherichia coli MG1655 4.64 Mb

E. coli 16 3.26 Mb

Lambda homologous recombination and P1 transduction

1377.2 kb (29.7%)

Much slower growth rate, longer and wider cell shape than the parental strain, abnormal nucleoid organisation

N/A

17

Escherichia coli MG1655 4.64 Mb

E. coli 33a 2.83 Mb

Homologous recombination and FLP-FRT site-specific recombination and P1 transduction

1806.6 kb (38.9%)

Sensitive to oxidative stress

N/A

18

Bacillus subtilis 168 4.22 Mb

B. subtilis Δ6 3.9 Mb

Homologous recombination and excision

320 kb (7.7%)

Normal growth, intracellular carbon metabolism, competence and sporulation, but changed motility

No significant difference of α-amylase (AmyQ) secretion compared to the wild-type

10

Bacillus subtilis 168 4.22 Mb

B. subtilis PG10 2.76 Mb and PS38 2.68 Mb

Homologous recombination and counter selection

PG10 1.46 Mb (36%) PS38 1.54 Mb (36.5%)

Comparable growth to the wild- type in complex medium

PG10 showed substantially higher capacity for protein secretion

20,54

Streptomyces avermitilis 9.02 Mb

S. avermitilis SUKA 7.35–7.51 Mb

Homologous recombination and Cre-mediated recombination

1.52–1.67 Mb (16.9–18.54%)

Could grow on the minimum medium

Higher streptomycin and cephamycin C production than their native hosts; produce the plant terpenoid Intermediate, amorpha-4,11-diene, successfully

12

Schizosaccharomyces pombe 12.09 Mb

S. pombe A8 11.43 Mb

Homologous recombination by the Latour method

657.3 Kb (5.43%)

Decreased uptake of glucose and some amino acids compared with the parental strain

2.7-fold increase in the concentration of cellular adenosine triphosphate; 1.7- and 1.8-fold increase in the production of green fluorescent protein and secreted human growth hormone, respectively

13