Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation

Abstract

Background The production and processing of animal-based products generates many collagen-rich by-products, which have received attention both for exploitation to increase their added value and to reduce their negative environmental impact. The collagen-rich by-products can be hydrolyzed by collagenases for further utilization. Therefore, collagenases are of benefit for efficient collagen materials processing. An alternative and safe way to produce secreted collagenases is needed. Results Two collagenases from Hathewaya histolytica, ColG and ColH, were successfully secreted by the yeast Saccharomyces cerevisiae. Compared with the native signal peptide of collagenase, the α-factor leader is more efficient in guiding collagenase secretion. Collagenase secretion was significantly increased in YPD medium by supplementing with calcium and zinc ions. Recombinant collagenase titers reached 68 U/mL and 55 U/mL for ColG and ColH, respectively. Collagenase expression imposed metabolic perturbations on yeast cells; substrate consumption, metabolites production and intracellular cofactor levels changed in engineered strains. Both recombinant collagenases from yeast could hydrolyze soluble and insoluble collagen materials. Recombinant ColG and ColH showed a synergistic effect on efficient collagen digestion. Conclusions Sufficient calcium and zinc ions are essential for active collagenase production by yeast. Collagenase secretion was increased by optimization of expression cassettes. Collagenase expression imposed metabolic burden and cofactor perturbations on yeast cells, which could be improved through metabolic engineering. Our work provides a useful way to produce collagenases for collagen resource utilization. Graphical Abstract

CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori

“…Moreover, these cuticular protein genes had covered almost all the cuticular protein families of RR-1, RR-2, Tweedle, CPFL, CPG, and CPH, with the exception of RR-3 [ 25 ]. Intriguingly, another group of molecular structure such as collagens were all down-regulated in mutation of BmMETTL4 ( Figure 7 C), whereas collagenase responsible for degrading native collagen were significantly up-regulated ( Figure 7 D) [ 26 , 27 ]. These analyses indicated important roles of structural proteins in the process of embryogenesis.…”

“…Alternatively, it was found that there were 7 collagenases highly expressed in BmMETTL4 mutant. Collagenases are enzymes with serine hydrolase activity, which are capable of degrading native collagen under physiological conditions [ 27 ]. The significant up-regulation of these enzymes would further destroy the molecular structure activity of collagens.…”

Optimization of Protein Folding for Improved Secretion of Human Serum Albumin Fusion Proteins in Saccharomyces cerevisiae

“…The expression of amylase-HSA fusion protein disturbed the intracellular redox environment of S. cerevisiae strains, necessitating the production of more byproducts to balance redox cofactors and maintain cell homeostasis. − …”

Correction: Secretion of collagenases by Saccharomyces cerevisiae for collagen degradation

“…Following publication of the original article [ 1 ], the author noticed an error in Additional file 1: Fig. S5 within the additional information.…”