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A Novel Lipase as Aquafeed Additive for Warm-Water Aquaculture.

Ran C, He S, Yang Y, Huang L, Zhou Z - PLoS ONE (2015)

Bottom Line: Results showed that supplementation of LipG1 significantly improved the gut and heptaopancreas lipase activity of fish fed with palm oil diet.Consistently, improved feed conversion ratio and growth performance were recorded in the LipG1 feeding group, to levels comparable to the group of fish fed with soybean oil diet.Collectively, LipG1 exhibited good potential as an aquafeed additive enzyme, and deserves further characterization as the representative of a novel group of lipases.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P. R. China.

ABSTRACT
A novel Acinetobacter lipase gene lipG1 was cloned from DNA extracted from intestinal sample of common carp (Cyprinus carpio), and expressed in E. coli BL21. The encoded protein was 406 amino acids in length. Phylogenetic analysis indicated that LipG1 and its relatives comprised a novel group of true lipases produced by Gram-negative bacteria. LipG1 showed maximal activity at 40℃ and pH 8.0 when pNP decanoate (C10) was used as the substrate, and remained high activity between 20℃ and 35℃. Activity of the lipase was promoted by Ca2+ and Mg2+, and inhibited by Zn2+ and Cu2+. Moreover, LipG1 is stable with proteases, most commercial detergents and organic solvents. Substrate specificity test indicated that LipG1 can hydrolyse pNP esters with acyl chain length from C2 to C16, with preference for medium-chain pNP esters (C8, C10). Lastly, LipG1 was evaluated as an aquafeed additive for juvenile common carp (Cyprinus carpio). Results showed that supplementation of LipG1 significantly improved the gut and heptaopancreas lipase activity of fish fed with palm oil diet. Consistently, improved feed conversion ratio and growth performance were recorded in the LipG1 feeding group, to levels comparable to the group of fish fed with soybean oil diet. Collectively, LipG1 exhibited good potential as an aquafeed additive enzyme, and deserves further characterization as the representative of a novel group of lipases.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic tree of true lipases produced by Gram-negative bacteria, including LipG1 and its relatives.Abbreviations and sequence accession numbers: PaPAO1 (P. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; LipG1, this study; Ab348935 (A. baumannii 348935), WP_034704719.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Pflu (P. fluorescens SIK W1), BAA02012.1; Smar (Serratia marcescens), BAA02519.1.
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pone.0132049.g004: Phylogenetic tree of true lipases produced by Gram-negative bacteria, including LipG1 and its relatives.Abbreviations and sequence accession numbers: PaPAO1 (P. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; LipG1, this study; Ab348935 (A. baumannii 348935), WP_034704719.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Pflu (P. fluorescens SIK W1), BAA02012.1; Smar (Serratia marcescens), BAA02519.1.

Mentions: Bacterial true lipases comprise the family I of bacterial lipolyitc enzymes, and were further classified into six subfamilies [25]. True lipases from Gram-negative bacteria were assigned into the first three subfamilies. Subfamily I.1 were lipases from Pseudomonas aeruginosa, Vibrio cholerae, Acinetobacter calcoaceticus, etc. Subfamily I.2 mainly comprises lipases produced by Burkholderia, while I.3 contains enzymes from two distinct species: Ps. fluorescens and Serratia marcescens [25]. Many lipase genes have been cloned from Acinetobacter [14, 15, 17, 18, 19]. The Acinetobacter lipases were assigned to subfamily I.1 of bacterial true lipases [25], and were proposed as a separate ‘Acinetobacter’ clade based on similarity of amino acid sequences [15, 26]. However, amino acid sequence alignment showed that LipG1 isn’t homologous to any of the above mentioned Acinetobacter lipases and doesn’t belong to the typical ‘Acinotebacter’ lipase clade. Based on the phylogenetic tree predicted from multiple sequence alignment of the lipases (Fig 4), and considering the deviation in the position of the catalytic residue His (Fig 1), we propose that LipG1 and its relatives comprise another subfamily of true lipases from Gram-negative bacteria. The lipases homologous with LipG1 in the BLASTp list all originated from genomic sequence analysis rather than experimental characterization. Also, around half of the hits with over 60% identity in the BLASTp list were designated as hypothetical proteins from Acinetobacter strains. Therefore, LipG1 represents a novel group of lipases that deserves further characterization.


A Novel Lipase as Aquafeed Additive for Warm-Water Aquaculture.

Ran C, He S, Yang Y, Huang L, Zhou Z - PLoS ONE (2015)

Phylogenetic tree of true lipases produced by Gram-negative bacteria, including LipG1 and its relatives.Abbreviations and sequence accession numbers: PaPAO1 (P. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; LipG1, this study; Ab348935 (A. baumannii 348935), WP_034704719.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Pflu (P. fluorescens SIK W1), BAA02012.1; Smar (Serratia marcescens), BAA02519.1.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4492967&req=5

pone.0132049.g004: Phylogenetic tree of true lipases produced by Gram-negative bacteria, including LipG1 and its relatives.Abbreviations and sequence accession numbers: PaPAO1 (P. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; LipG1, this study; Ab348935 (A. baumannii 348935), WP_034704719.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Pflu (P. fluorescens SIK W1), BAA02012.1; Smar (Serratia marcescens), BAA02519.1.
Mentions: Bacterial true lipases comprise the family I of bacterial lipolyitc enzymes, and were further classified into six subfamilies [25]. True lipases from Gram-negative bacteria were assigned into the first three subfamilies. Subfamily I.1 were lipases from Pseudomonas aeruginosa, Vibrio cholerae, Acinetobacter calcoaceticus, etc. Subfamily I.2 mainly comprises lipases produced by Burkholderia, while I.3 contains enzymes from two distinct species: Ps. fluorescens and Serratia marcescens [25]. Many lipase genes have been cloned from Acinetobacter [14, 15, 17, 18, 19]. The Acinetobacter lipases were assigned to subfamily I.1 of bacterial true lipases [25], and were proposed as a separate ‘Acinetobacter’ clade based on similarity of amino acid sequences [15, 26]. However, amino acid sequence alignment showed that LipG1 isn’t homologous to any of the above mentioned Acinetobacter lipases and doesn’t belong to the typical ‘Acinotebacter’ lipase clade. Based on the phylogenetic tree predicted from multiple sequence alignment of the lipases (Fig 4), and considering the deviation in the position of the catalytic residue His (Fig 1), we propose that LipG1 and its relatives comprise another subfamily of true lipases from Gram-negative bacteria. The lipases homologous with LipG1 in the BLASTp list all originated from genomic sequence analysis rather than experimental characterization. Also, around half of the hits with over 60% identity in the BLASTp list were designated as hypothetical proteins from Acinetobacter strains. Therefore, LipG1 represents a novel group of lipases that deserves further characterization.

Bottom Line: Results showed that supplementation of LipG1 significantly improved the gut and heptaopancreas lipase activity of fish fed with palm oil diet.Consistently, improved feed conversion ratio and growth performance were recorded in the LipG1 feeding group, to levels comparable to the group of fish fed with soybean oil diet.Collectively, LipG1 exhibited good potential as an aquafeed additive enzyme, and deserves further characterization as the representative of a novel group of lipases.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P. R. China.

ABSTRACT
A novel Acinetobacter lipase gene lipG1 was cloned from DNA extracted from intestinal sample of common carp (Cyprinus carpio), and expressed in E. coli BL21. The encoded protein was 406 amino acids in length. Phylogenetic analysis indicated that LipG1 and its relatives comprised a novel group of true lipases produced by Gram-negative bacteria. LipG1 showed maximal activity at 40℃ and pH 8.0 when pNP decanoate (C10) was used as the substrate, and remained high activity between 20℃ and 35℃. Activity of the lipase was promoted by Ca2+ and Mg2+, and inhibited by Zn2+ and Cu2+. Moreover, LipG1 is stable with proteases, most commercial detergents and organic solvents. Substrate specificity test indicated that LipG1 can hydrolyse pNP esters with acyl chain length from C2 to C16, with preference for medium-chain pNP esters (C8, C10). Lastly, LipG1 was evaluated as an aquafeed additive for juvenile common carp (Cyprinus carpio). Results showed that supplementation of LipG1 significantly improved the gut and heptaopancreas lipase activity of fish fed with palm oil diet. Consistently, improved feed conversion ratio and growth performance were recorded in the LipG1 feeding group, to levels comparable to the group of fish fed with soybean oil diet. Collectively, LipG1 exhibited good potential as an aquafeed additive enzyme, and deserves further characterization as the representative of a novel group of lipases.

No MeSH data available.


Related in: MedlinePlus