Retinal production from β-carotene by β-carotene 15,15′-dioxygenase from an unculturable marine bacterium

Abstract

The conversion of beta-carotene to retinal by a recombinant beta-carotene 15,15'-dioxygenase (Blh protein) from an unculturable marine bacterium was optimized in aqueous solution. Toluene was optimal solvent for the dissolution of beta-carotene and the optimal solution for the conversion reaction contained 2.4% (w/v) Tween 20, 0.15 U enzyme/ml, and 350 mg beta-carotene/l. Under these conditions, the enzyme produced 181 mg retinal/l after 20 h. This is the highest reported value for the retinal concentration from beta-carotene.

Production of β-apo-10′-carotenal from β-carotene by human β-carotene-9′,10′-oxygenase expressed in E. coli

“…When the concentration of Tween 40 was varied from 0.6 to 6.0% (w/ v), the highest concentration of b-apo-10 0 -carotenal occurred at 2.4% (w/v) (data not shown). These results are similar to those obtained in retinal production by BCO1s (Kim et al 2007(Kim et al , 2008(Kim et al , 2010.…”

“…The data represent the means and standard deviations from three separate experiments Biotechnol Lett (2011) 33:1195-1200 1197 micelles might have prevented contact between the enzyme at the outside surface of the micelle and the b-carotene located in the hydrophobic core (Shikama 2006), thereby decreasing b-apo-10 0 -carotenal production at b-carotene concentrations above 300 mg/l. The optimal concentrations of b-carotene for retinal production using BCO1s from mice (Kim et al 2007), humans (Kim et al 2008), and marine bacteria (Kim et al 2010) were reported to be 200, 200, and 350 mg/l, respectively.c b-Apo-10 0 -carotenal production from b-carotene by human BCO2 under the optimum conditions…”

Purification of β‐carotene 15,15′‐monooxygenase from pig intestine and its enzymatic hydrolysis of pigment in soybean oil

“…The result is the same as that of Kim's study (Kim et al ., ). For recombinant mouse β‐carotene 15,15′‐monooxygenase, under its optimal condition (5.0% (w/v) Tween 40, 0.20 U/mL enzyme, 200 mg β‐carotene/L), the enzyme produced 72 mg retinal/L after 20 h, with a conversation yield of 36% (w/w), which was higher than the β‐carotene conversion rate of our study (Kim et al ., ). It may be that they use Escherichia coli as the host of the β‐carotene 15,15′‐monooxygenase, and the recombinant protein production system in E. coli has a commercial advantage, thereby increasing the yield of retinol.…”

“…Therefore, increasing the amount of Tween added has no significant effect on increasing the production of vitamin A. According to Kim et al, Tween 20 (at 2.4%, w/v) gave the highest retinal production (Kim et al, 2010).…”

“…High-performance liquid chromatography (HPLC) analyses of vitamin A were performed based on the method of Kim, with slight modification (Kim et al, 2010). Vitamin A was detected using an ODS-A column (50 9 3 mm).…”