Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum

Wördemann, Ramona; Wiefel, Lars; Wendisch, Volker F.; Steinbüchel, Alexander

doi:10.1186/s13568-021-01217-5

AMB Express

Table 3 Comparison of different C. glutamicum strains for CGP synthesis

From: Incorporation of alternative amino acids into cyanophycin by different cyanophycin synthetases heterologously expressed in Corynebacterium glutamicum

Strains	DW [g/l]	Cyanophycin
		Solubility	CGP content [%DW]	Amino acid composition[mol%] *
		Solubility	CGP content [%DW]	Asp	Arg	Lys	Glu	Orn	Cit
ATCC 13032
pVWEx1::cphA₆₃₀₈Δ1	15.9 ± 0.1	Insoluble	1.9 ± 0.5	50.1	38.9	8.7	0.5	1.0	0.8
pVWEx1::cphA₆₃₀₈Δ1	15.9 ± 0.1	Soluble	4.0 ± 0.1	51.9	26.9	10.9	1.2	6.6	2.5
pVWEx1::cphA₆₃₀₈Δ1_C595S	15.6 ± 0.2	Insoluble	1.4 ± 0.3	54.7	30.0	13.6	0.3	0.8	0.6
pVWEx1::cphA₆₃₀₈Δ1_C595S	15.6 ± 0.2	Soluble	8.3 ± 2.9	51.3	27.5	10.7	0.9	6.4	3.2
pVWEx1::cphA_Dh	16.6 ± 0.4	Insoluble	0.0 ± 0.0	–	–	–	–	–	–
pVWEx1::cphA_Dh	16.6 ± 0.4	Soluble	36.2 ± 0.7	49.3	40.1	6.9	2.3	0.6	0.8
DM1279
pVWEx1::cphA₆₃₀₈Δ1	11.6 ± 0.3	Insoluble	12.2 ± 2.1	52.9	30.3	16.2	0.2	0.0	0.4
pVWEx1::cphA₆₃₀₈Δ1	11.6 ± 0.3	Soluble	2.9 ± 0.6	45.6	21.0	29.7	1.0	2.2	0.5
pVWEx1::cphA₆₃₀₈Δ1_C595S	11.7 ± 0.2	Insoluble	13.1 ± 2.3	48.0	29.8	21.6	0.2	0.0	0.4
pVWEx1::cphA₆₃₀₈Δ1_C595S	11.7 ± 0.2	Soluble	2.1 ± 0.5	42.7	19.8	33.5	1.0	2.6	0.4
pVWEx1::cphA_Dh	14.4 ± 0.6	Insoluble	0.0 ± 0.0	–	–	–	–	–	–
pVWEx1::cphA_Dh	14.4 ± 0.6	Soluble	24.2 ± 1.9	48.3	33.3	15.4	2.2	0.8	0.0
ORN2(P_gdh4)
pVWEx1::cphA₆₃₀₈Δ1	11.6 ± 0.2	Insoluble	1.9 ± 0.2	47.6	42.7	0.0	0.3	9.1	0.3
pVWEx1::cphA₆₃₀₈Δ1	11.6 ± 0.2	Soluble	3.2 ± 0.7	46.0	43.5	0.0	0.6	9.4	0.5
pVWEx1::cphA₆₃₀₈Δ1_C595S	11.7 ± 0.1	Insoluble	2.4 ± 0.2	42.8	47.9	0.0	0.3	8.7	0.3
pVWEx1::cphA₆₃₀₈Δ1_C595S	11.7 ± 0.1	Soluble	2.3 ± 0.7	41.7	44.2	0.0	0.7	12.6	0.8
pVWEx1::cphA_Dh	12.9 ± 0.3	Insoluble	0.0 ± 0.0	–	–	–	–	–	–
pVWEx1::cphA_Dh	12.9 ± 0.3	Soluble	18.6 ± 1.6	42.8	43.7	0.0	1.4	11.7	0.4

The strains were cultivated for 72 h in 100 ml CGXII medium in a 500 ml Erlenmeyer flask with two baffles at 130 rpm and 30 °C. The cultures were inoculated with an OD₆₀₀ of about 0.2 and after 4 h IPTG was added. For the ORN2(P_gdh4) strains an additional 15 mM arginine was added to the medium. The cells dry mass was determined, and the contained cyanophycin was isolated and analysed. The values indicate the mean and standard deviation of at least three independent cultivations
^* The values have been rounded to one decimal place

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