Difference between revisions of "Os09g0567800"
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===Expression=== | ===Expression=== | ||
| − | + | Lipases are important biocatalysts showing many interesting properties with industrial applications. Previously, different isoforms of lipases, Lipase-I and Lipase-II from rice (Oryza sativa) have been purified and characterized. Lipase-II identified as the major lipase in rice bran is designated as rice bran lipase (RBL). An exploration of expression in four different E. coli expression systems analyzed: BL21(DE3)pLysS, RIL(DE3)pLysS, Rosetta(DE3)pLysS and Origami(DE3)pLysS indicated that E. coli was not a suitable host. Expression with supplement of rare codons in Rosetta (DE3)pLysS and RIL(DE3)pLysS resulted in highest expression as insoluble inclusion bodies. The hurdles of expression in E. coli were overcome by expression as a secretory protein in P. pastoris X-33. The expression of lipase in shake flasks was optimized to achieve the maximum recombinant lipase activity of 152.6 U/mL. The purified recombinant lipase had a specific activity of 998 U/mg toward triacetin. The pH and temperature optimum of native and recombinant enzymes were pH 7.4 and 25 ± 2 °C, respectively. Both the lipases showed higher activity toward short chain triacylglycerol and unsaturated fatty acid enriched oils. Computational modeling and molecular docking studies reveal that the catalytic efficiency of the lipase correlates with the distance of the nucleophilic Ser(175)-OH and the scissile ester bond. The shorter the distance, the greater is the turnover of the substrate.[4] | |
===Evolution=== | ===Evolution=== | ||
Revision as of 04:28, 6 June 2014
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Contents
Annotated Information
Function
This gene encodes an lipolytic enzyme(lipase) involved in the degradation of fat,which will form fatty acids, glycerol and monoglyceride or diacylglycerol.[1]lipolytic enzyme perform essential roles in the digestion, transport and processing of dietary lipids (e.g. triglycerides, fats, oils) in most, if not all, living organisms.Most lipases act at a specific position on the glycerol backbone of lipid substrate (A1, A2 or A3)(small intestine). Lipases are involved in diverse biological processes ranging from routine metabolism of dietary triglycerides to cell signaling[2] and inflammation.[3] Thus, some lipase activities are confined to specific compartments within cells while others work in extracellular spaces.Other lipase enzymes, such as pancreatic lipases, are secreted into extracellular spaces where they serve to process dietary lipids into more simple forms that can be more easily absorbed and transported throughout the body.As biological membranes are integral to living cells and are largely composed of phospholipids, lipases also play important roles in cell biology.
Expression
Lipases are important biocatalysts showing many interesting properties with industrial applications. Previously, different isoforms of lipases, Lipase-I and Lipase-II from rice (Oryza sativa) have been purified and characterized. Lipase-II identified as the major lipase in rice bran is designated as rice bran lipase (RBL). An exploration of expression in four different E. coli expression systems analyzed: BL21(DE3)pLysS, RIL(DE3)pLysS, Rosetta(DE3)pLysS and Origami(DE3)pLysS indicated that E. coli was not a suitable host. Expression with supplement of rare codons in Rosetta (DE3)pLysS and RIL(DE3)pLysS resulted in highest expression as insoluble inclusion bodies. The hurdles of expression in E. coli were overcome by expression as a secretory protein in P. pastoris X-33. The expression of lipase in shake flasks was optimized to achieve the maximum recombinant lipase activity of 152.6 U/mL. The purified recombinant lipase had a specific activity of 998 U/mg toward triacetin. The pH and temperature optimum of native and recombinant enzymes were pH 7.4 and 25 ± 2 °C, respectively. Both the lipases showed higher activity toward short chain triacylglycerol and unsaturated fatty acid enriched oils. Computational modeling and molecular docking studies reveal that the catalytic efficiency of the lipase correlates with the distance of the nucleophilic Ser(175)-OH and the scissile ester bond. The shorter the distance, the greater is the turnover of the substrate.[4]
Evolution
Please input evolution information here.
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Labs working on this gene
Department of Biochemistry and Molecular Biology ,Oklahoma University Health Sciences Center, Oklahoma City, USA
References
1.Chi-Sun Wang, Jean A. Hartsuck. (1993)Bile salt-activated lipase. A multiple function lipolytic enzyme.Biochimica et Biophysics Acta, 1166: l-19
2.Spiegel S, Foster D, and R Kolesnick.(1996) "Signal transduction through lipid second messengers". Current Opinion in Cell Biology 8(2): 159–67.
3.Tjoelker LW, Eberhardt C, Unger J, Trong HL, Zimmerman GA, McIntyre TM, Stafforini DM, Prescott SM, and PW Gray.(1995) "Plasma platelet-activating factor acetylhydrolase is a secreted phospholipase A2 with a catalytic triad". J Biol Chem 270(43): 25481–7.
4.Vijayakumar KR, Gowda LR.(2013)Rice (Oryza sativa) lipase: molecular cloning, functional expression and substrate specificity.Protein Expr Purif.88(1):67-79.
Structured Information
| Gene Name |
Os09g0567800 |
|---|---|
| Description |
Lipolytic enzyme, G-D-S-L family protein |
| Version |
NM_001070500.2 GI:297609976 GeneID:4347890 |
| Length |
4001 bp |
| Definition |
Oryza sativa Japonica Group Os09g0567800, complete gene. |
| Source |
Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group
Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP
clade; Ehrhartoideae; Oryzeae; Oryza.
|
| Chromosome | |
| Location |
Chromosome 9:23503081..23507081 |
| Sequence Coding Region |
23503402..23503914,23503998..23504260,23506420..23506695,23506817..23506913 |
| Expression | |
| Genome Context |
<gbrowseImage1> name=NC_008402:23503081..23507081 source=RiceChromosome09 preset=GeneLocation </gbrowseImage1> |
| Gene Structure |
<gbrowseImage2> name=NC_008402:23503081..23507081 source=RiceChromosome09 preset=GeneLocation </gbrowseImage2> |
| Coding Sequence |
<cdnaseq>atggaggaggagagcagcacgctcatacggcgccggactagtttgccggcgaccacggtgatgatggtggtcctcctcctcctcctctgctgtaactgtggtgtcgaggtggtggtggctacggctgatgagtcgtctccggcgccggtcggcaaaggaggaagtgatcatggcagttgccccggcggcgacggcgacggcgaagggtacaggaagcagctgtgggtgtttggcgactcgtacgcggacacgggaaacctgggtaaccttgggagggagctgacccacgcctggtactacccctacggcatcaccttcccgcgacaccccaccggccgcttctccgacggccgcgtcctcaccgacttcgttgcttcggccgttggcatcgcgacgccggtggcgtacaagctgcggcggcgcggcgggcacggcggcgaggtggcgtcgcgcgggatgaacttcgcggtgggcgggtcgggcgtgctggacacgggctacttccagcgcaacatcagctcgcagatcgacctgttccagaagcagctgcgcggctgcggccccaccggcgtcgcgctcgtcgtcgtctccggcaacgactactccgccgtcgtagacaagaacaacggcacaagcgaggcggcgatcgcgtacatcccgacggtggtgagggggctgcgggagcaactccggcggctgcgcgacgaggtagggatgaagaaggtggtggtgaccaacctccaccccatgggctgcaccccgtacttcacgcggctgctcaactactccggctgcgacacgctggccaacgccgggtccgaccagcacaacgccgccctccgctccgtcctccacgacctcgaccccgccaacaccaccttcctcctcctcgacctccacacccccttcctcaacctcatcaccgccgccgccgacgacaagttcccggtgcggctgcggccgtgctgcgagacgttcacggcggacggccactgcgggcaggaggacgaagccggcaacaagcagtacacggtgtgcgacgacccggagcggcacttctactgggacgacgtgcaccccacgcaggccgcctgggccgccgtcgcccaagccttcacccccgccatccaccgcttcctctccacctga</cdnaseq> |
| Protein Sequence |
<aaseq>MEEESSTLIRRRTSLPATTVMMVVLLLLLCCNCGVEVVVATADE SSPAPVGKGGSDHGSCPGGDGDGEGYRKQLWVFGDSYADTGNLGNLGRELTHAWYYPY GITFPRHPTGRFSDGRVLTDFVASAVGIATPVAYKLRRRGGHGGEVASRGMNFAVGGS GVLDTGYFQRNISSQIDLFQKQLRGCGPTGVALVVVSGNDYSAVVDKNNGTSEAAIAY IPTVVRGLREQLRRLRDEVGMKKVVVTNLHPMGCTPYFTRLLNYSGCDTLANAGSDQH NAALRSVLHDLDPANTTFLLLDLHTPFLNLITAAADDKFPVRLRPCCETFTADGHCGQ EDEAGNKQYTVCDDPERHFYWDDVHPTQAAWAAVAQAFTPAIHRFLST</aaseq> |
| Gene Sequence |
<dnaseqindica>3168..3680#2822..3084#387..662#169..265#gatcgcgccaccaccccctctcctctcacactatcaattgctcttgtcatctctgatctgcctgccgctgcgccagctgcctcctccgatctctctctcactctcacacacacgtactagctagcaagtgcacacagacaatgtgactctgtgagagatctatcgggcatggaggaggagagcagcacgctcatacggcgccggactagtttgccggcgaccacggtgatgatggtggtcctcctcctcctcctctgctgtaactgtaagctgcatgctctgccctccatatctgctcccccacttttactagtattaaattctagctatctactactcaaattcatttactgctgaaacctcaactcccataatgtgacgtataggtggtgtcgaggtggtggtggctacggctgatgagtcgtctccggcgccggtcggcaaaggaggaagtgatcatggcagttgccccggcggcgacggcgacggcgaagggtacaggaagcagctgtgggtgtttggcgactcgtacgcggacacgggaaacctgggtaaccttgggagggagctgacccacgcctggtactacccctacggcatcaccttcccgcgacaccccaccggccgcttctccgacggccgcgtcctcaccgacttcgttggtacgcccacgcctccgccatctctctctatatatcttcatgctatgctaattaattactcctactgtcatacgtactgtacatgtcatgctacattatatcgatctgtacattctcttggtcatgcatattacagcgccatatcccatatatcgtacctacgaccaaaattaaactaaacgctctctgaaaaaccaatcaacaaaatacatacaacctcatgttcatgtaggggtataaactataattaatgttcatacaaggagtatcactatcgatcaagctgctgctatgctaggtggaggatggcaatattgcattgatgtcgcccgggttcgttgggccagtggctgctaaccgaatttaattccatcctccgctgctgctgcgccagcccggccagcctcccaagaccaaagtgaacatatggccctgtctagttccttaaaaaacttttcgcaaaaacattatatcaaatctctgtacataggtatagaatattaaatatagataaaaagaaaaactaattgcacagtttatatgtaaatcacgaaatgaatcttttaaacctaattaatccatgattagtcataagtgctacaacaacccacatgtgctaataacggcttaattaggctcaaaagattcttctcgcggtttccatacgagttatgaaattagtttttttcattcgtgtccgaaaaccctttccgacatatgatcaaacatccgatgtgacatctaaaattttcatttcatcaactaaacaggcccatatatgtactggagtacgtgccttttatacagacagactactactagtcactcattttacaacctgcacaatggaaatttaccactcaaaatttttcataggcagactgtacatcattttcgtttctaaccaaaatatatataggtgatcacaaaaatcacaatgataggcaaataggagtaactaactaaaagaagaatctttgcattatattggctctttttcgcggatggaaattgactcagttcatcgcatctggataattcattcatcattcattcattctttgtttgttttttcatgccgacttaactgaccgcacatgcataaaagcctcaggctgcagctagtccatgcctgcactgcatagtgacgacgtctgtattattctctttctttttacctaccctataataggtctccaaattagcacgccctgtcctattgttactgttattgctctcactcctcgcttaattaggcttgacatgtacatgtccaaaacataaaacacatttatcaatcgaaccgacattttggttgcaaatctagtgaacaactaagcaagttatggtgaggcgtcacaatcaactgtaccctgcccttcaaaacaataactaaggctgtgtttttcagcgcaaagtttggattttggttgaaattgaagatgatgtgattgaaaagttgtgtgtgtatgacaggttgatgtgatggaaaaggactgaagtttggatccaaactttagatctaaacacagcctaactgtatatcagtaacagtcctgtcgcgcgccgtttctgaagagatgatatatagatcctctgtgtttaatttgcattcagcatttatcagttttatctaattaattgttgtcttaaaagatgtactaaaattaaactggctagtatttgatgaactctctctttttctatacaagtagcacttgtttttagaggtaagttccttcctagtgttcttcagactagtttaagaggttgagcagcattcatgtttagttgcagcctagctattggaggcaaactaaactaaaagctgaattcgctctcacctgctctactgacgtgcgtatgttccataccgcatctcatgtgccggcagtattttatttatttttaatcactctctcaacaactagtaaaagctaatgtatcctgcacatttgtagagagtaattaaacagcacgcgcgcgtgcgtgcatctatcatgtcagataatgtgccatacaattatctggctgaaagctaaccttaaccattaaagtattttgcagtgtttatttgtttgtttttataacagagataatagtaataaggttttaactggtacgtagtatggttgaatttaaaatattggcgggtgcagcttcggccgttggcatcgcgacgccggtggcgtacaagctgcggcggcgcggcgggcacggcggcgaggtggcgtcgcgcgggatgaacttcgcggtgggcgggtcgggcgtgctggacacgggctacttccagcgcaacatcagctcgcagatcgacctgttccagaagcagctgcgcggctgcggccccaccggcgtcgcgctcgtcgtcgtctccggcaacgactactccgccgtcgtagacaagaacaacggcacaagcgtaagctgcaattgcaatacaaccacaccggccatctcgatctatatatcatatctcctctctgaacatgcatgtatgaacaggaggcggcgatcgcgtacatcccgacggtggtgagggggctgcgggagcaactccggcggctgcgcgacgaggtagggatgaagaaggtggtggtgaccaacctccaccccatgggctgcaccccgtacttcacgcggctgctcaactactccggctgcgacacgctggccaacgccgggtccgaccagcacaacgccgccctccgctccgtcctccacgacctcgaccccgccaacaccaccttcctcctcctcgacctccacacccccttcctcaacctcatcaccgccgccgccgacgacaagttcccggtgcggctgcggccgtgctgcgagacgttcacggcggacggccactgcgggcaggaggacgaagccggcaacaagcagtacacggtgtgcgacgacccggagcggcacttctactgggacgacgtgcaccccacgcaggccgcctgggccgccgtcgcccaagccttcacccccgccatccaccgcttcctctccacctgaacgaacgaatgaatactcaccaaccagctttctcatttccgacgaaattccgcgaaatttcgacgtttctatatggctcggaattaagtttcgattcgaatttcgaaggttaaacagtagatttgatcgaaattcagtagatttgctccttgcacttcttcttcttcttcttcttcgtcttcttgaacaactttttttggggcctatctaatttattattacctttggatgatgatttgttgcaactggatagcagtattactgatttttttttttgagcaaagggcgagagccctccatttccattaatagaaatgaaaa</dnaseqindica> |
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