Abstract
Dietary supplements and functional foods are becoming increasingly popular complements to regular diets. A recurring ingredient is the essential cofactor vitamin B12 (B12). Microalgae are making their way into the dietary supplement and functional food market but do not produce B12, and their B12 content is very variable. In this study, the suitability of using the human B12-binding protein intrinsic factor (IF) to enrich bioavailable B12 using microalgae was tested. The IF protein was successfully expressed from the nuclear genome of the model microalga Chlamydomonas reinhardtii and the addition of an N-terminal ARS2 signal peptide resulted in efficient IF secretion to the medium. Co-abundance of B12 and the secreted IF suggests the algal produced IF protein is functional and B12-binding. Utilizing IF expression could be an efficient tool to generate B12-enriched microalgae in a controlled manner that is suitable for vegetarians and, potentially, more bioavailable for humans.
Highlights
There is an increasing awareness of the effect of diet on human health and, in particular, in the development of illness and chronic disease
Constructs for nuclear transformation of a wall-deficient C. reinhardtii strain (CC-849) were assembled with the human gene encoding the mature sequence of the B12 -binding protein intrinsic factor (IF) flanked by 50 and 30 untranslated regions (UTRs) of the PSAD gene
The ble marker downstream of the gene of interest on the pCrEX1 plasmid was used to select for positive transformants after transformation by agitation of a DNA-cell mixture with glass beads
Summary
There is an increasing awareness of the effect of diet on human health and, in particular, in the development of illness and chronic disease. This has led to an upward trend in consumer demand for naturally occurring bioactives that can be delivered in the form of dietary supplements or incorporated into functional foods. Vitamin B12 or cobalamin (B12 ), is an essential cofactor produced by certain prokaryotes via two main complex biosynthetic pathways [1]. B12 is an essential cofactor required for a range of cellular metabolism functions (e.g., DNA synthesis and methylation, mitochondrial metabolism) [2].
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