Table 1 Strains and plasmids used in this study

Strains

BSGN5

B. subtilis 168 derivate, ΔnagPΔgamPΔgamAΔnagAΔnagBΔldh::lox72

[30]

BSGN10

B. subtilis 168 derivate, ΔnagPΔgamPΔgamAΔnagAΔnagBΔldhΔalsRSD::lox72

[5]

BSGN12

B. subtilis 168 derivate, ΔnagPΔgamPΔgamAΔnagAΔnagBΔldhΔptaΔackA::lox72, ΔalsRSD::Pveg-ΔermC+14/7A-ecglmS

[6]

BSGN12-P_veg-urease

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_veg

This study

BSGN12-P_xylA-urease

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_xylA

This study

BSGN12-P_abrB-urease

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_abrB

This study

BSGN13

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_hag

This study

BSGN12-P_ffh-urease

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_ffh

This study

BSGN12-P_licH-urease

BSGN12 derivate, expression of urease from Bacillus paralicheniformis under the control of promoter P_licH

This study

Plasmids

This study

p7Z6

pMD18-T containing lox71-zeo-lox66 cassette

[19]

pTSC

Em^rAmp^r; temperature sensitive in B. subtilis

[19]

pP₄₃-cMyc (M-Rm)-CeGNA1

key enzyme CeGNA1 expressing vector

[6]

pP₄₃-cMyc (M-Rm)-CeGNA1-Q155V/C158G

pP₄₃-cMyc (M-Rm)-CeGNA1 derivate, with 155Gln and 158Cys of CeGNA1 mutated to 155Val and 158Gly, respectively

This study

pP₄₃-6His-CeGNA1

pP₄₃NMK derivate with CeGNA1 cloned

[6]

pCold-CeGNA1

pCold III derivate, containing wild type CeGNA1, with His tag fused to the N terminal

This study

pCold-CeGNA1-Q155V/C158G

pCold III derivate, containing mutant CeGNA1-Q155V/C158G, with His tag fused to the N terminal

This study

pP_vegEcGlmS-1

pUC57-Amp derivate, containing the synthesized expression cassette, terminator I-P_veg-TSS-RBS₀-terminator II

[6]

pStop1622

Amp^r, Tet^r, E. coli-B. megaterium shuttle vector

[31]