| Lactase |
| Identifiers |
| EC number |
3.2.1.108 |
| CAS number |
9031-11-2 |
| Databases |
| IntEnz |
IntEnz view |
| BRENDA |
BRENDA entry |
| ExPASy |
NiceZyme view |
| KEGG |
KEGG entry |
| MetaCyc |
metabolic pathway |
| PRIAM |
profile |
| PDB structures |
RCSB PDB PDBe PDBsum |
| Gene Ontology |
AmiGO / EGO |
| Search |
| PMC |
articles |
| PubMed |
articles |
|
Glycosylceramidase
(Phlorizin hydrolase) |
| Identifiers |
| EC number |
3.2.1.62 |
| CAS number |
9033-10-7 |
| Databases |
| IntEnz |
IntEnz view |
| BRENDA |
BRENDA entry |
| ExPASy |
NiceZyme view |
| KEGG |
KEGG entry |
| MetaCyc |
metabolic pathway |
| PRIAM |
profile |
| PDB structures |
RCSB PDB PDBe PDBsum |
| Gene Ontology |
AmiGO / EGO |
| Search |
| PMC |
articles |
| PubMed |
articles |
|
| Lactase |
| Identifiers |
| Symbol |
LCT |
| Alt. symbols |
LAC; LPH; LPH1 |
| Entrez |
3938 |
| HUGO |
6530 |
| OMIM |
603202 |
| RefSeq |
NM_002299 |
| UniProt |
P09848 |
| Other data |
| EC number |
3.2.1.108 |
| Locus |
Chr. 2 q21 |
Lactase (also known as lactase-phlorizin hydrolase, or LPH), a part of the β-galactosidase family of enzymes, is a glycoside hydrolase involved in the hydrolysis of the disaccharide lactose into constituent galactose and glucose monomers. Lactase is present predominantly along the brush border membrane of the differentiated enterocytes lining the villi of the small intestine.[1] In humans, lactase is encoded by the LCT gene.[2][3]
Lactase is essential for digestive hydrolysis of lactose in milk. Deficiency of the enzyme causes lactose intolerance.[4]
The optimum temperature for lactase is about 77 °F (25 °C) for its activity[5] and has an optimum pH of 6.[1]
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Contents
- 1 Function and mechanism
- 2 Structure and biosynthesis
- 3 Genetic expression and regulation
- 4 Industrial use
- 5 See also
- 6 References
- 7 External links
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Function and mechanism
Lactase can hydrolyze a variety of substrates. While it is most notably a member of the β-galactosidase enzymatic class, lactase also has glucosidase and glycosylceramidase activity.[6] In metabolism, the β-glycosidic bond in D-lactose is hydrolyzed to form D-galactose and D-glucose, which can be absorbed through the intestinal walls and into the bloodstream. The overall reaction that lactase catalyzes is C12H22O11 + H2O → C6H12O6 + C6H12O6 + heat. Lactase also catalyzes the conversion of phlorizin to phloretin and glucose.
The catalytic mechanism of D-lactose hydrolysis retains the substrate anomeric configuration in the products.[7] While the details of the mechanism are uncertain, the stereochemical retention is achieved through a double displacement reaction. Studies of E. coli lactase have proposed that hydrolysis is initiated when a glutamate nucleophile on the enzyme attacks from the axial side of the galactosyl carbon in the β-glycosidic bond.[8] The removal of the D-glucose leaving group may be facilitated by Mg-dependent acid catalysis.[8] The enzyme is liberated from the α-galactosyl moiety upon equatorial nucleophilic attack by water, which produces D-galactose.[7]
Substrate modification studies have demonstrated that the 3’-OH and 2’-OH moieties on the galactopyranose ring are essential for enzymatic recognition and hydrolysis.[6] The 3’-hydroxy group is involved in initial binding to the substrate while the 2’- group is not necessary for recognition but needed in subsequent steps. This is demonstrated by the fact that a 2-deoxy analog is an effective competitive inhibitor (Ki = 10mM).[6] Elimination of specific hydroxyl groups on the glucopyranose moiety does not completely eliminate catalysis.[6]
Proposed mechanism of lactose hydrolysis.
Structure and biosynthesis
Pre-pro-lactase, the primary translation product, has a single polypeptide primary structure consisting of 1927 amino acids.[2] It can be divided into five domains: (i) a 19 amino acid cleaved signal sequence; (ii) a large prosequence domain that is not present in mature lactase; (iii) the mature lactase segment; (iv) a membrane spanning hydrophobic anchor; and (v) a short hydrophilic carboxyl terminus.[2] The signal sequence is cleaved in the endoplasmic reticulum, and the resulting 215 kDa pro-LPH is sent to the Golgi, where it is heavily glycosylated and proteolytically processed to its mature form.[9] The prodomain has been shown to act as an intramolecular chaperone in the ER, preventing trypsin cleavage and allowing LPH to adopt the necessary 3-D structure to be transported to the Golgi apparatus.[10]
Schematic of processing and localization of human lactase translational product.
Mature human lactase consists of a single 160 kDa polypeptide chain that localizes to the brush border membrane of intestinal epithelial cells. It is oriented with the N-terminus outside the cell and the C-terminus in the cytosol.[2] LPH contains two catalytic glutamic acid sites. In the human enzyme, the lactase activity has been connected to Glu-1749 while Glu-1273 is the site of phlorizin hydrolase function.[11]
Genetic expression and regulation
Lactase is encoded by a single genetic locus on chromosome 2.[12] It is expressed exclusively by mammalian small intestine enterocytes and in very low levels in the colon during fetal development.[12] Humans are born with high levels of lactase expression. In most of the world’s population, lactase transcription is down-regulated after weaning, resulting in diminished lactase expression in the small intestine.[12] Diminished lactase expression causes the common symptoms of adult-type hypolactasia, or lactose intolerance.
Some population segments exhibit lactase persistence resulting from a mutation that is postulated to have occurred 5000-10,000 years ago, coinciding with the rise of cattle domestication.[13] This mutation has allowed almost half of the world’s population to metabolize lactose without symptoms. Studies have linked the occurrence of lactase persistence to two different single-nucleotide polymorphisms about 14 and 22 kilobases upstream of the 5’-end of the LPH gene.[14] Both mutations, C→T at position -13910 and T→ A at position -22018, have been independently linked to lactase persistence.[15]
The lactase promoter is 150 base pairs long and is located just upstream of the site of transcription initiation.[15] The sequence is highly conserved in mammals, suggesting that critical cis-transcriptional regulators are located nearby.[15] Cdx-2, HNF-1α, and GATA have been identified as transcription factors.[15] Studies of hypolactasia onset have demonstrated that despite polymorphisms there is little difference in lactase expression in infants, showing that the mutations become increasingly relevant during development.[16] Ιt is hypothesized that developmentally-regulated DNA-binding proteins down-regulate transcription or destabilize mRNA transcripts, causing decreased LPH expression after weaning.[16]
Industrial use
Lactase produced commercially can be extracted both from yeasts such as Kluyveromyces fragilis and Kluyveromyces lactis and from fungi, such as Aspergillus niger and Aspergillus oryzae.[17] Its primary commercial use is to break down lactose in milk to make it suitable for people with lactose intolerance. However, the U.S. Food and Drug Administration (FDA) has not formally evaluated the effectiveness of these products.[18] Lactase is also used in the manufacture of ice cream. Because glucose and galactose are sweeter than lactose, lactase produces a sweeter ice cream. Lactose also crystallizes at the low temperatures of ice cream; however, its constituent products, glucose and galactose, remain dissolved and contribute to a smoother texture. Lactase is used in the conversion of whey into syrup.
Lactase is also used to screen for blue white colonies in the MCS of various plasmid vectors in Escherichia coli or other bacteria.[citation needed]
See also
- MCM6
- Lactose
- Developmental regulation of lactase expression in mammals
References
- ^ a b Skovbjerg H, Sjöström H, Norén O (March 1981). "Purification and characterisation of amphiphilic lactase/phlorizin hydrolase from human small intestine". Eur. J. Biochem. 114 (3): 653–61. doi:10.1111/j.1432-1033.1981.tb05193.x. PMID 6786877.
- ^ a b c d Mantei N, Villa M, Enzler T, Wacker H, Boll W, James P, Hunziker W, Semenza G (September 1988). "Complete primary structure of human and rabbit lactase-phlorizin hydrolase: implications for biosynthesis, membrane anchoring and evolution of the enzyme". EMBO J. 7 (9): 2705–13. . PMID 2460343. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=457059.
- ^ Harvey CB, Fox MF, Jeggo PA, Mantei N, Povey S, Swallow DM (July 1993). "Regional localization of the lactase-phlorizin hydrolase gene, LCT, to chromosome 2q21". Ann. Hum. Genet. 57 (Pt 3): 179–85. doi:10.1111/j.1469-1809.1993.tb01593.x. PMID 8257087.
- ^ Järvelä I, Torniainen S, Kolho KL (2009). "Molecular genetics of human lactase deficiencies". Ann. Med. 41 (8): 568–75. doi:10.1080/07853890903121033. PMID 19639477.
- ^ Hermida C, Corrales G, Cañada FJ, Aragón JJ, Fernández-Mayoralas A (July 2007). "Optimizing the enzymatic synthesis of beta-D-galactopyranosyl-D-xyloses for their use in the evaluation of lactase activity in vivo". Bioorg. Med. Chem. 15 (14): 4836–40. doi:10.1016/j.bmc.2007.04.067. PMID 17512743.
- ^ a b c d Fernandez P, Cañada F, Jiménez-Barbero J, Martín-Lomas, M (July 1995). "Substrate specificity of small-intestinal lactase: Study of the steric effects and hydrogen bonds involved in enzyme-substrate interaction". Carbohydr. Res. 271 (1): 31–42. doi:10.1016/0008-6215(95)00034-Q. PMID 7648581.
- ^ a b Sinnott M (November 1990). "Catalytic mechanisms of enzymic glycosyl transfer". Chem. Rev. 90 (7): 1171–1202. doi:10.1021/cr00105a006.
- ^ a b Juers D, Heightman T, Vasella A, McCarter J, Mackenzie L, Withers S, Matthews B (December 2001). "A structural view of the action of Escherichia coli (lacZ) β-galactosidase". Biochemistry 40 (49): 14781–94. doi:10.1021/bi011727i. PMID 11732897.
- ^ Naim HY, Sterchi E, Lentze M (January 1987). "Biosynthesis and maturation of lactase-phlorizin hydrolase in the human small intestinal epithelial cells". Biochem. J. 241 (2): 427–34. . PMID 3109375. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1147578.
- ^ Naim HY, Jacob R, Naim H, Sambrook J, Gething M (October 1994). "The pro region of human intestinal lactase-phlorizin hydrolase". J. Biol. Chem. 269 (43): 26933–43. PMID 7523415.
- ^ Zecca L, Mesonero J, Stutz A, Poirée J, Guidicelli J, Cursio R, Gloor S, Semenza G (September 1998). "Intestinal lactase-phlorizin hydrolase (LPH): the two catalytic sites; the role of the pancreas in pro-LPH maturation". FEBS Lett. 435 (2–3): 225–8. doi:10.1016/S0014-5793(98)01076-X. PMID 9762914.
- ^ a b c Troelsen J, Mitchelmore C, Spodsberg N, Jensen A, Norén O, Sjöström H (March 1997). "Regulation of lactase-phlorizin hydrolase gene expression by the caudal-related homoeodomain protein Cdx-2". Biochem. J. 322 (Pt. 3): 833–838. . PMID 9148757. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1218263.
- ^ Bersaglier T, Sabeti P, Patterson N, Vanderploeg T, Schaffner S, Drake J, Rhodes M, Reich D, Hirschhorn J (June 2004). "Genetic Signatures of Strong Recent Positive Selection at the Lactase Gene". Am. J. Hum. Gen. 74 (6): 1111–20. doi:10.1086/421051. . PMID 15114531. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1182075.
- ^ Kuokkanen M, Enattah N, Oksanen A, Savilhahtl E, Orpana A, Järvela I (May 2003). "Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia". Gut. 52 (5): 647–52. doi:10.1136/gut.52.5.647. . PMID 12692047. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=1773659.
- ^ a b c d Troelsen J (May 2005). "Adult-type hypolactasia and regulation of lactase expression". Biochim Biophys Acta. 1723 (1–3): 19–32. doi:10.1016/j.bbagen.2005.02.003. PMID 15777735.
- ^ a b Wang Y, Harvey C, Hollox E, Phillips A, Poulter M, Clay P, Walker-Smith J, Swallow D (June 1998). "The genetically programmed down-regulation of lactase in children". Gastroenterology 114 (6): 1230–6. doi:10.1016/S0016-5085(98)70429-9. PMID 9609760.
- ^ Seyis I, Aksoz N (2004). "Production of lactase by Trichoderma sp". Food Technol Biotechnol 42: 121–124. http://public.carnet.hr/ftbrfd/42-121.pdf.
- ^ Tarantino, LM (2003-12-03). "Agency Response Letter GRAS Notice No. GRN 000132". U.S. Food and Drug Administration. http://www.fda.gov/Food/FoodIngredientsPackaging/GenerallyRecognizedasSafeGRAS/GRASListings/ucm153949.htm. Retrieved 2009-09-21.
External links
- The Lactase Protein
- E. coli β-galactosidase: PDB 1JYY
- The use of lactases in the dairy industry
- The Recent Evolution of Lactose Tolerance
- Gene Ontology for Lactase
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Hydrolase: sugar hydrolases (EC 3.2)
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| 3.2.1: Glycoside hydrolases |
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Disaccharidase
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Sucrase/Sucrase-isomaltase/Invertase · Maltase · Trehalase · Lactase
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Glucosidases
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Cellulase · Alpha-glucosidase (Acid, Neutral AB, Neutral C) · Beta-glucosidase (cytosolic) · Debranching enzyme
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Other
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Amylase (Alpha-Amylase) · Chitinase · Lysozyme · Neuraminidase (NEU1, NEU2, NEU3, NEU4, Bacterial neuraminidase, Viral neuraminidase) · Galactosidases (Alpha, Beta) · alpha-Mannosidase · Glucuronidase · Hyaluronidase · Pullulanase · Glucosylceramidase (lysosomal, non-lysosomal) · Galactosylceramidase · Alpha-N-acetylgalactosaminidase (NAGA) · Alpha-N-acetylglucosaminidase · Fucosidase · Hexosaminidase (HEXA, HEXB) · Iduronidase · Maltase-glucoamylase · Heparanase (HPSE2)
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3.2.2: Hydrolysing
N-Glycosyl compounds |
DNA glycosylases: Oxoguanine glycosylase
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- B
- enzm
- 1.1
- 2
- 3
- 4
- 5
- 6
- 7
- 8
- 10
- 11
- 13
- 14
- 15-18
- 2.1
- 2.7.10
- 2.7.11-12
- 3.1
- 3.1.3.48
- 3.4.21
- 4.1
- 5.1
- 6.1-3
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Metabolism: carbohydrate metabolism · fructose and galactose enzymes
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| Fructose / Fructolysis |
Hepatic fructokinase · Aldolase B · Triokinase
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| Sorbitol |
Sorbitol dehydrogenase · Aldose reductase
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| Galactose / Galactolysis |
Galactokinase · Galactose-1-phosphate uridylyltransferase/UDP-glucose 4-epimerase
Aldose reductase
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| Lactose |
Lactose synthase · Lactase
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| Mannose |
Mannose phosphate isomerase
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mt, k, c/g/r/p/y/i, f/h/s/l/o/e, a/u, n, m
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k, cgrp/y/i, f/h/s/l/o/e, au, n, m, epon
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m(A16/C10),i(k, c/g/r/p/y/i, f/h/s/o/e, a/u, n, m)
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