Enzymatic preparation of food‐grade l‐α‐glycerylphosphorylcholine from soy phosphatidylcholine or fractionated soy lecithin

Abstract

L-α-Glycerylphosphorylcholine (L-α-GPC) is a biosynthetic precursor for the neurotransmitter acetylcholine in humans, making it a useful as a cognitive enhancer for treating patients with stroke and dementia, including Alzheimer's disease. The aim of this study was to prepare L-α-GPC via Novozym 435 (an immobilized Candida antarctica lipase B)-catalyzed hydrolysis of soy phosphatidylcholine or a fractionated soy lecithin, from which triacylglycerols were completely removed, followed by foodgrade solvent extraction of L-α-GPC from the reaction products. The reaction was performed in n-hexane-water biphasic media in a stirred-batch reactor. Phosphatidylcholine was completely hydrolyzed to L-α-GPC under optimal conditions: temperature, 55 C; water content, 100 wt% of the substrate weight; enzyme loading, 10 wt % of the substrate weight; and reaction time of 6 hr (for soy phosphatidylcholine) or 8 hr (for fractionated soy lecithin). Water-soluble fractions of the reaction products containing 98.6 area% L-α-GPC (from soy phosphatidylcholine) or 52.4 area% glycerophosphodiesters, including L-α-GPC (from fractionated soy lecithin), were obtained after phase separation of the media. The resulting products would be suitable for use as food-grade cognitive enhancers because of the use of enzymatic reaction and food-grade solvent extraction. K E Y W O R D Sfood-grade cognitive enhancer, lecithin, L-α-glycerylphosphorylcholine, Novozym 435, phosphatidylcholine

Immobilized Phospholipase A1-Catalyzed Preparation of l-α-Glycerylphosphorylcholine from Phosphatidylcholine

“…26 We previously found that Novozym 435, a commercially available immobilized form of CALB, was an effective biocatalyst for the preparation of L-α-GPC from soy PC using a similar reaction system to that employed in this study. 17 Novozym 435 is manufactured by adsorbing CALB on Lewatit VP OC 1600. In our previous work, soy PC was completely hydrolyzed to L-α-GPC within 6 h. The catalytic activity of CALB immobilized on Duolite A568, Amberlite XAD7HP, Accurel MP1000, and Lewatit VP OC 1600 was evaluated for the hydrolysis of soy PC to L-α-GPC, similarly to LU.…”

“…The activity of the Lewatit VP OC 1600-immobilized LU prepared in this study was lower than that of Novozym 435 using a similar reaction system. 17 Using a similar reaction system, Bang et al 6 found that free LU could completely hydrolyze soy PC to L-α-GPC after 30 h, with the longer reaction time related to a lower E n (13 mg) and lower water composition in the n-hexane−water biphasic media.…”

“…Recently, we demonstrated that enzymatic hydrolysis can completely convert soy PC to l -α-GPC in n -hexane–water biphasic media. , The reaction was performed using a commercial immobilized lipase, Novozym 435 (Candida antarctica lipase B, immobilized on macroporous acrylic resin), or LU (free form) as the biocatalyst. Using the immobilized lipase, l -α-GPC was easily recovered in very high purity (∼99 g/100 g) after filtration of the enzyme followed by simple phase separation of the biphasic media.…”

Value-Added Products from Ethanol Fermentation—A Review

“…The use of toxic substrates can produce α-GPC that is not safe for use in food and, thus, not marketable. Alternatively, α-GPC has been produced by enzymatic hydrolysis of PC in aqueous media [52,57,[64][65][66], employing phospholipases (Figure 2) [64]. Enzymatic production of α-GPC is advantageous, as the amounts of chemical reagents can be reduced, thereby making a comparably inexpensive product that is suited for use in food and cosmetic products [64].…”

“…Alternatively, α-GPC has been produced by enzymatic hydrolysis of PC in aqueous media [52,57,[64][65][66], employing phospholipases (Figure 2) [64]. Enzymatic production of α-GPC is advantageous, as the amounts of chemical reagents can be reduced, thereby making a comparably inexpensive product that is suited for use in food and cosmetic products [64]. However, enzymatic production of α-GPC can also be difficult due to the limited solubility of PC in aqueous phases and long reaction times (e.g., low activity) for phospholipases [67][68][69].…”