N-terminal truncation of PhaCBP-M-CPF4 and its effect on PHA production

Table 3 PHA production of the C. necator PHB⁻4 transformants harbouring the full-length phaC_BP-M-CPF4 or truncated phaC_BP-M-CPF4 using fructose or CPKO with its molecular weights

Carbon source	Strain	Cell dry weight (g/L)	PHA content (wt%)	Residual biomass (g/L)	PHA composition (mol%)		Molecular weights (× 10⁵ Da)
Carbon source	Strain	Cell dry weight (g/L)	PHA content (wt%)	Residual biomass (g/L)	3HB	3HHx	M_n	M_w	PDI
Fructose	PHB⁻4/ pBBR1-C_BP-M-CPF4	8.3 ± 0.1^v	78.0 ± 2.9^v	1.8 ± 0.2^vw	100 ± 0^v	0 ± 0^v	9.8 ± 0.5^v	22.8 ± 0.1^v	2.3 ± 0.1^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 G8	8.4 ± 0.2^v	81.9 ± 4.1^v	1.5 ± 0.3^v	100 ± 0^v	0 ± 0^v	11.2 ± 0.5^w	26.1 ± 0.5^w	2.3 ± 0.1^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 A27	4.6 ± 0.1^w	56.5 ± 2.9^w	2.0 ± 0.2^vw	100 ± 0^v	0 ± 0^v	10.9 ± 0.8^v	26.9 ± 0.3^w	2.5 ± 0.5^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 T74	2.4 ± 0.0^x	17.8 ± 0.4^x	2.0 ± 0.0^w	100 ± 0^v	0 ± 0^v	6.7 ± 0.1^x	18.2 ± 0.1^x	2.7 ± 0.0^w
	PHB⁻4/pBBR1-C_BP-M-CPF4 D104	1.8 ± 0.0^y	0.5 ± 0.1^y	1.8 ± 0.0^v	100 ± 0^v	0 ± 0^v	ND	ND	ND
CPKO	PHB⁻4/ pBBR1-C_BP-M-CPF4	11.3 ± 0.9^v	75.8 ± 2.4^v	2.7 ± 0.5^v	94 ± 0^v	6 ± 0^v	3.7 ± 0.3^v	9.3 ± 0.4^v	2.5 ± 0.1^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 G8	10.2 ± 0.3^v	75.9 ± 2.7^v	2.5 ± 0.3^v	95 ± 0^w	5 ± 0^w	4.6 ± 0.1^w	11.2 ± 0.2^w	2.4 ± 0.0^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 A27	7.5 ± 0.4^x	59.7 ± 5.1^w	3.0 ± 0.4^v	96 ± 0^x	4 ± 0^x	5.7 ± 0.2^x	14.1 ± 0.1^x	2.5 ± 0.1^v
	PHB⁻4/pBBR1-C_BP-M-CPF4 T74	3.8 ± 0.1^y	54.1 ± 5.4^w	1.7 ± 0.1^w	97 ± 0^y	3 ± 0^y	5.3 ± 0.1^y	10.5 ± 0.0^y	2.0 ± 0.0^w
	PHB⁻4/pBBR1-C_BP-M-CPF4 D104	1.8 ± 0.3^z	10.3 ± 2.7^x	1.6 ± 0.2^w	99 ± 0^z	1 ± 0^z	ND	ND	ND

The bacterial cells were cultivated in 50 mL of MM supplemented with 0.054% (w/v) of urea as nitrogen source, 50 µg/mL kanamycin, 2% (v/v) of fructose or 1% (v/v) of CPKO as carbon source. The bacterial cells were shaken at 200 rpm for 48 h at 30 °C. The values reported are in means ± standard deviations of triplicate. The superscripts v, w, x, y, and z represent the significant differences in the GC and GPC data using one-way ANOVA (p < 0.05). Abbreviations: 3HB, 3-hydroxybutyrate; 3HHx, 3-hydroxyhexanoate; ND, not determined

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