Biocatalyst for the production of 25-OH-Vitamin D3 and other derivatives by recombinant Bacillus megaterium strains

Background
The biotechnological production of 25-OH Vitamin D3 is of great importance, as this inactivated position 25 within vitamin D3 is hardly achievable by means of organic synthesis, especially in terms of a regio- and stereoselective manner.
Within the human body, 25-OH vitamin D3 is converted into the biological active form of vitamin D3 which is important for the Ca2+ deposit into bones. Furthermore, in case of hepatic cirrhosis, its substitution is essential. It has anti-carcinogenic effects and acts protective in terms of autoimmune and cardiovascular diseases.
25-OH vitamin D3 is also used as a standard in clinical laboratory diagnostics. Altogether, 25-OH vitamin D3 is valuable concerning biotechnology, pharmacy and medicine.

Invention
Biochemists of Saarland University identified and optimized two cytochrome P450 systems which are capable of performing a regio- and stereoselective hydroxylation of vitamin D3 in one step, transferring vitamin D3 to 25-OH and other hydroxyl- derivatives of vitamin D3. The enzymes belong to the CYP109 family.
One of these enzymes is capable of synthesizing 25-OH vitamin D3 as a major product and three side products in vitro producing a yield of 26 mg/l/24h.
The second one gives rise to 25-OH vitamin D3 as the only product producing a yield of 5 mg/l/24h.

Advantages
• Synthesis of 25-OH vitamin D3 and other hydroxyl-derivatives in high yields

o 25-OH vitamin D3 and three side products→ 26 mg/l/24h
o 25-OH vitamin D3 → 5 mg/l/24h

• Low production costs

• No toxic side products as in classical organic synthesis

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Universität des Saarlandes Wissens- und Technologietransfer GmbH PatentVerwertungsAgentur der saarländischen Hochschulen
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Ansprechpartner
Dr. Conny Clausen, Dr. Nicole Comtesse, Dr. Frank Döbrich, Dr. Susanne Heiligenstein, Dr. Christof Schäfer

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