Abstract
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.
Highlights
Bile salt-stimulated lipase (BSSL)[1], which is known as carboxyl ester lipase (CEL), bile salt-dependent lipase (BSDL) or carboxyl ester hydrolase (CEH), is secreted from the exocrine pancreas into the intestinal lumen in all species examined to date[2, 3]
When comparing recombinant human BSSL (rhBSSL) purified from transgenic milk with native BSSL purified from human milk and rhBSSL expressed by Chinese hamster ovary (CHO) cells we found no notable differences in physicochemical and other characteristics
A Neo selection marker was downstream of BSSL. Human lactoferrin bacterial artificial chromosome (BAC) (hLF BAC) have a 90-kb 5’UTR and a 30-kb 3’UTR, and they were used as transcriptional regulating elements
Summary
Bile salt-stimulated lipase (BSSL)[1], which is known as carboxyl ester lipase (CEL), bile salt-dependent lipase (BSDL) or carboxyl ester hydrolase (CEH), is secreted from the exocrine pancreas into the intestinal lumen in all species examined to date[2, 3]. It is secreted by the mammary gland in some species, such as humans[4], but not cows or goats[5].
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