Microbial Production of Retinyl Palmitate and Its Application as a Cosmeceutical

Abstract

Chemically synthesized retinyl palmitate has been widely used in the cosmetic and biotechnology industry. In this study, we aimed to demonstrate the microbial production of retinyl palmitate and the benefits of microbial retinyl palmitate in skin physiology. A heterologous retinyl palmitate biosynthesis pathway was reconstructed in metabolically engineered Escherichia coli using synthetic expression modules from Pantoea agglomerans, Salinibacter ruber, and Homo sapiens. High production of retinyl palmitate (69.96 ± 2.64 mg/L) was obtained using a fed-batch fermentation process. Moreover, application of purified microbial retinyl palmitate to human foreskin HS68 fibroblasts led to increased cellular retinoic acid-binding protein 2 (CRABP2) mRNA level [1.7-fold (p = 0.001) at 100 μg/mL], acceleration of cell proliferation, and enhancement of procollagen synthesis [111% (p < 0.05) at 100 μg/mL], strongly indicating an anti-ageing-related effect of this substance. These results would pave the way for large-scale production of retinyl palmitate in microbial systems and represent the first evidence for the application of microbial retinyl palmitate as a cosmeceutical.

Microbial Production of Bioactive Retinoic Acid Using Metabolically Engineered Escherichia coli

“…Collectively, the blhSR and raldhHS genes were functionally expressed in the BETA-1/pSTVM-blh-raldh strain, generating retinoic acid (0.52 ± 0.10 mg/L) from β-carotene. Notably, the presence of retinol (peak 2) and retinal (peak 3) in the extract of BETA-1/pSTVM2-blh was the same as that in our previous study [11].…”

“…The blh SR gene encoding β-carotene 15,15′-oxygenase (BlhSR) from S. ruber , which was previously cloned in the plasmid pUCMr-blh [ 11 ], was utilized, and the raldh HS gene encoding retinaldehyde dehydrogenase 2 (RALDH2, GenBank accession number, AB015226.1) from the Hep3B cell line was cloned into pUCMr to construct pUCMr-raldh ( Table 1 ). To construct the retinoic acid pathway, the two genes encoding BlhSR and RALDH2 were redesigned as an individual expression module, and then assembled into the plasmid pSTVM2, using the USER ® cloning method, resulting in pSTVM2-blh-raldh.…”

“…To enhance the expression levels of the two genes in E. coli, the 5 -untranslated regions (5 -UTRs) of the two genes were engineered by using 26-bp mRNA-stabilizing sequences, which were obtained by using UTR designer (http://sbi.postech.ac.kr/utr_designer, accessed on 7 May 2020). Three 5 -UTRs of blhSR (UTR12, UTR37, and UTR46), which were investigated in our previous study [11], were utilized as pUCMr12-blh, pUCMr37-blh, and pUCMr46-blh, respectively, in this study. In a similar way described in [11], two 5 -UTRs of RALDH2 (named UTRH for high expression, and UTRM for moderate expression) were predicted using UTR designer, and two plasmids (pUCMrH-raldh and pUCMrMraldh) were constructed.…”

“…Microbial production of RA has received attention as an alternative process for retinoic acid. Although retinol and retinyl ester have been produced in microbial host strains, including Although retinol and retinyl ester have been produced in microbial host strains, including Escherichia coli [9,10,11] and Saccharomyces cerevisiae [12], there are no reports on the microbial biosynthesis of retinoic acid. As E. coli has been exploited as a microbial host system for retinoid production, it can serve as a host for the redesign and reconstruction of the retinoic acid pathway.…”

Comparing Sediment Microbiomes in Contaminated and Pristine Wetlands along the Coast of Yucatan

“…In particular, the halophilic bacterium S . ruber has a retinal-based light-driven pump known as xanthorhodopsin [ 44 , 45 , 46 ]. Sesquiterpenoid and triterpenoid have commercial potential in the pharmaceutical sector due to their bioactivities such as antiviral, anticancer, anti-inflammatory, wound-healing properties, pigments, hormones and signaling molecules, antibiotics, antifeedants, or pollinator attractants.…”