Enhanced hyaluronic acid production in Bacillus subtilis by coexpressing bacterial hemoglobin.
Bacillus subtilis strains that can produce hyaluronic acid (HA) were constructed by integrating the HA synthase gene (hasA) and the UDP-glucose dehydrogenase gene of group C Streptococcus (hasB) or of B. subtilis itself (tauD) into the amyE locus of the B. subtilis chromosome. All of the inserted genes were under the control of a strong constitutive vegII promoter of B. subtilis. Although HA production could be achieved by expressing hasA alone, coexpressing hasB or tauD with hasA could enhance HA production at least 2-fold. To replenish the energy consumed for HA biosynthesis, Vitreoscilla hemoglobin (VHb) was coexpressed with the HA-expressing genes. With the expression of VHb, not only the cell concentration was enhanced 25%, but also HA production was further increased by 100%. About 1.8 g/L of HA was obtained by the recombinant strain B. subtilis carrying VHb, hasA, and tauD genes in the expression cassette after 30 h cultivation.
Chien
Liang-Jung
LJ
Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan.
Lee
Cheng-Kang
CK
D001412:Bacillus subtilis / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods*; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000302:isolation & purification / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D013291:Streptococcus / Q000378:metabolism*; D014169:Transformation, Bacterial / Q000502:physiology; D054793:Truncated Hemoglobins / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D006454:Hemoglobins; D011994:Recombinant Proteins; D054793:Truncated Hemoglobins; C082613:hemoglobin protein, Vitreoscilla; D006820:Hyaluronic Acid