Increased dipicolinic acid production with an enhanced <i>spoVF</i> operon in <i>Bacillus subtilis</i> and medium optimization

<div><p>Dipicolinic acid (DPA) is a multi-functional agent for cosmetics, antimicrobial products, detergents, and functional polymers. The aim of this study was to design a new method for producing DPA from renewable material. The <i>Bacillus subtilis spoVF</i> operon encodes enzymes for DPA synthase and the part of lysine biosynthetic pathway. However, DPA is only synthesized in the sporulation phase, so the productivity of DPA is low level. Here, we report that DPA synthase was expressed in vegetative cells, and DPA was produced in the culture medium by replacement of the <i>spoVFA</i> promoter with other highly expressed promoter in <i>B. subtilis</i> vegetative cells, such as <i>spoVG</i> promoter. DPA levels were increased in the culture medium of genetically modified strains. DPA productivity was significantly improved up to 29.14 g/L in 72 h culture by improving the medium composition using a two-step optimization technique with the Taguchi methodology.</p></div

MOESM2 of A novel GH10 xylanase from Penicillium sp. accelerates saccharification of alkaline-pretreated bagasse by an enzyme from recombinant Trichoderma reesei expressing Aspergillus β-glucosidase

Additional file 2: Figure S2. SDS-PAGE of enzyme preparation JN11H and JNK25H. SDS-PAGE was carried out with Any kD Mini-PROTEAN TGX Precast Protein Gels (Bio-Rad, Hercules, CA) and the gel was activated and imaged using the ChemiDoc MP imaging system (Bio-Rad).3 μg of protein were loaded on each gel. Lanes: M, Precision Plus Protein Unstained Standard; 1, JN11H enzyme preparation produced by T. reesei strain X3AB1; 2, JNK25H enzyme preparation produced by strain X2PX10. Cellobiohydrolases (CBH 1, CBH2), endoglucanases (EG1 and EG2), xylanases (XYN1 and XYN2) and β-xylosidases (BXL) from Trichoderma reesei, and heterologously expressed proteins (β-glucosidase AaBGL1 from Aspergillus aculeatus and xylanase PspXyn10 from Penicillium sp. KSM-F532) are shown

MOESM1 of A novel GH10 xylanase from Penicillium sp. accelerates saccharification of alkaline-pretreated bagasse by an enzyme from recombinant Trichoderma reesei expressing Aspergillus β-glucosidase

Additional file 1: Figure S1. Multiple alignments and phygogenetic tree of PspXyn10, PspXyn10 orthologs and TrXyn3. (a) Multiple alignments of the PspXyn10, PspXyn10 orthologs and TrXyn3. The amino acid sequence of PspXyn10 was compared with those of endo-1,4-beta-xylanase from Penicillium brasilianum (OOQ87260), endo-1,4-beta-xylanase D from Penicillium subrubescens (OKO90312), endo-1,4-beta-xylanase from Rasamsonia emersonii CBS 393.64 (XP_013330999), hypothetical protein PENANT_c053G02808 from Penicillium antarcticum (OQD79351), endo-1,4-beta-xylanase Xyl10A from Talaromyces cellulolyticus (GAM37231) and TrXyn3 from Trichoderma reesei (BAA89465) by multiple alignment. The alignment was created using ClustalW2 on Genetyx Version 12 software (Genetyx). Amino acid numbers are shown on the left and right. Black boxes indicate invariant residues. Gray boxes indicate the residues conserved in more than half of aligned sequences. (b) Phylogenetic tree of the PspXyn10, PspXyn10 orthologs and TrXyn3. Phylogenetic tree of the amino acid sequences were created using the NJ method under 1000 times bootstrap conditions using Genetyx Version 12 software (Genetyx)