Development of dried liposomes containing beta-galactosidase for the digestion of lactose in milk.
The hydrolyzed-lactose milk for lactase-deficient subjects has a sweeter taste than whole milk, and some subjects dislike its taste. In order to cope with this shortcoming, we examined whether beta-galactosidase, which hydrolyzes lactose, added to the whole milk in the form of dried liposomes, would be able to digest lactose in milk following the lysis of liposomes in the presence of bile salts. Dried liposomes containing beta-galactosidase were prepared in the presence of trehalose by the dehydration-rehydration vesicle method to overcome the instability of the conventional liposome suspension. The stability of liposomal membranes was evaluated by measuring the activity of entrapped beta-galactosidase under various storage conditions. By treating liposomes with trehalose, which was found to prevent the fusion of liposomes and the leakage of entrapped drug, the entrapping efficiency increased up to fourfold. Over 95% of dried liposomes which had been stored at 17 degrees C for 60 days were reconstituted to liposomes upon rehydration process. From the stability study, dried liposomes were found to retain 87% of beta-galactosidase activity at 17 degrees C after 60 days and to be more stable than the multilamellar vesicle suspension prepared without trehalose. The lysis study showed that dried liposomes were hardly lyzed in the simulated gastric fluid with pepsin, but lyzed immediately more than 90% in 0.01 M deoxycholic acid. Lactose hydrolysis in the presence of deoxycholic acid after the addition of dried liposome-entrapped beta-galactosidase to whole milk was proportional to the quantity of entrapped beta-galactosidase and the amount of dried liposomes added. These results demonstrate that beta-galactosidase entrapped in liposome is stable and reconstituted mostly upon rehydration, and can digest lactose in milk after the efficient lysis of liposomes in the presence of bile salts. This study implies that beta-galactosidase entrapped in liposome may be applied to whole milk for lactase-deficient subjects.
Kim
C K
CK
College of Pharmacy, Seoul National University, San 56-1, Shinlim-Dong, Kwanack-Gu, Seoul, South Korea. ckkim@plaza.snu.ac.kr
Chung
H S
HS
Lee
M K
MK
Choi
L N
LN
Kim
M H
MH
D000818:Animals; D001647:Bile Acids and Salts / Q000737:chemistry*; D003840:Deoxycholic Acid / Q000737:chemistry; D004355:Drug Stability; D005690:Galactose / Q000032:analysis; D007785:Lactose / Q000378:metabolism*; D008081:Liposomes / Q000138:chemical synthesis* / Q000737:chemistry; D008892:Milk / Q000737:chemistry / Q000378:metabolism*; D009994:Osmolar Concentration; D010434:Pepsin A / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000032:analysis; D013997:Time Factors; D014199:Trehalose / Q000737:chemistry; D001616:beta-Galactosidase / Q000378:metabolism / Q000494:pharmacology*
D001647:Bile Acids and Salts; D008081:Liposomes; D010713:Phosphatidylcholines; D003840:Deoxycholic Acid; D014199:Trehalose; D001616:beta-Galactosidase; D010434:Pepsin A; D007785:Lactose; D005690:Galactose