Purification and characterization of recombinant human bile salt-stimulated lipase expressed in milk of transgenic cloned cows

• Published in: PloS one 2017-05, Vol.12 (5), p.e0176864-e0176864
• Main Authors: Wang, Yuhang, Ding, Fangrong, Wang, Tao, Liu, Wenjie, Lindquist, Susanne, Hernell, Olle, Wang, Jianwu, Li, Jing, Li, Ling, Zhao, Yaofeng, Dai, Yunping, Li, Ning
• Format: Article
• Language: English
• Subjects: Absorption; Affinity; Affinity chromatography; Age; Agronomy; Amino acids; Animal genetic engineering; Animal models; Animal welfare; Animals; Animals, Genetically Modified; Antiinfectives and antibacterials; Attachment; Baby foods; Bile; Bile salts; Biochemistry; Biocompatibility; Biology and Life Sciences; Bioreactors; Biotechnology; Blood circulation; Blotting, Western; Breast feeding; Breast milk; Buffers; Cadmium; Catalysis; Catalytic activity; Cattle; Cell adhesion; Cholesterol; Cholic acid; Clinical trials; Cloning; Cloning, Organism; Cow's milk; Cows (Cattle); Dairy cattle; DC-SIGN protein; Differentiation; Digestion; Drinking water; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Esters; Fatty acids; Fetuses; Formulations; Genetic aspects; Genetic Vectors; Health aspects; Histochemistry; HIV; Hot Temperature; Human immunodeficiency virus; Humans; Hydrogen-Ion Concentration; Hydroxyapatite; Inactivation; Infant formulas; Infants; Intestine; Kinetics; Lipase; Lipase - chemistry; Lipase - genetics; Lipase - isolation & purification; Lipase - therapeutic use; Malabsorption Syndromes - drug therapy; Mammary gland; Medicine and Health Sciences; Mice; Milk; Pancreas; Pasteurization; Physiological aspects; Polymerase Chain Reaction; Premature infants; Proteins; Purification; Research and Analysis Methods; Rodents; Salt; Salts; Small for gestational age; Stomach; Surgical implants; Velocity; Viruses; Weight reduction
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0176864

Bile salt-stimulated lipase (BSSL) is a lipolytic digestive enzyme with broad substrate specificity secreted from exocrine pancreas into the intestinal lumen in all species and from the lactating mammary gland into the milk of some species, notably humans but not cows. BSSL in breast milk facilitates digestion and absorption of milk fat and promotes growth of small for gestational age preterm infants. Thus, purified recombinant human BSSL (rhBSSL) can be used for treatment of patients with fat malabsorption and expressing rhBSSL in the milk of transgenic cloned cows would therefore be a mean to meet a medical need. In the present study, a vector pBAC-hLF-hBSSL was constructed, which efficiently expressed active rhBSSL in milk of transgenic cloned cows to a concentration of 9.8 mg/ml. The rhBSSL purified from cow milk had the same enzymatic activity, N-terminal amino acid sequence, amino acid composition and isoelectric point and similar physicochemical characteristics as human native BSSL. Our study supports the use of transgenic cattle for the cost-competitive, large-scale production of therapeutic rhBSSL.