Engineering Halomonas species TD01 for enhanced polyhydroxyalkanoates synthesis via CRISPRi

Table 1 Shake flask PHBV production by recombinants Halomonas sp. TD01 controlling prpC gene expression via CRISPRi

Recombinant TD01 strains	CDW (g/L)	PHBV (wt%)	3HV (mol%)
TD01	13.22 ± 0.20	76.08 ± 2.81	0.83 ± 0.08
TD-sgRNA	13.95 ± 0.71	74.73 ± 2.25	1.29 ± 0.47
TD-prpC1	14.99 ± 0.30	72.14 ± 2.37	1.79 ± 0.02
TD-prpC2	14.43 ± 0.45	75.06 ± 3.20	4.78 ± 0.83
TD-prpC3	14.08 ± 1.26	73.96 ± 7.18	5.72 ± 0.30
TD-prpC4	13.58 ± 0.57	80.14 ± 8.62	6.44 ± 0.43
TD-prpC5	13.59 ± 0.51	80.12 ± 3.27	8.16 ± 0.31
TD-prpC6	14.17 ± 0.23	82.20 ± 3.82	11.94 ± 0.80
TD-prpC7	13.54 ± 0.31	76.74 ± 0.80	12.15 ± 0.31
TD-prpC6prpC7	14.67 ± 0.78	73.78 ± 4.82	12.70 ± 0.27

The recombinants harboring CRISPRi system were cultivated in MM medium containing 30 g/L glucose and 1 g/L propionic acid at 37 °C for 48 h as described in “Methods”. CDW, cell dry weight; PHBV (wt%), the weight percent of PHBV in CDW; TD01, Halomonas sp. TD wild type strain; TD-sgRNA, TD01 strain harboring the pli-dCas9-sgRNA plasmid without any DNA target site; TD-prpC1, TD-prpC2, TD-prpC3, TD-prpC4, TD-prpC5, TD-prpC6, TD-prpC7, TD-prpC6prpC7, TD01 strains harboring the pli-dCas9-sgRNA plasmid with different sgRNA targets of gene prpC, respectively. All data were the average of three independent studies with standard deviations. Mean ± SE (n = 3)

ISSN: 1475-2859