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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

Alignment of amino acid sequences of representative lipases from subfamilies I.1 and I.2 of bacterial lipases and the group of lipases represented by LipG1.Abbreviations and sequence accession numbers: AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; AbBD5 (A. baumannii BD5), ABW70205.1; AcRAG-1 (A. calcoaceticus RAG-1), AAD29441.1; PaPAO1 (Ps. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; VcE1 (V. cholerae E1), P15493.2; PfIFO (P. fragi IFO-12049), P08658.2; PrvK80 (Proteus vulgaris K80), AAB01071.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Ab348935 (A. baumannii 348935), WP_034704719.1; LipG1, this study; Agyl (A. gyllenbergii), WP_032860322.1; Ajun (A. junii), WP_005402474.1. Symbols: ● Catalytic triad residues, ▲ Asp residues involved in calcium binding, ★ HG dipeptide of the oxyanion loop, ○ Putative triad residue, △Putative calcium binding Asp residues.
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pone.0132049.g001: Alignment of amino acid sequences of representative lipases from subfamilies I.1 and I.2 of bacterial lipases and the group of lipases represented by LipG1.Abbreviations and sequence accession numbers: AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; AbBD5 (A. baumannii BD5), ABW70205.1; AcRAG-1 (A. calcoaceticus RAG-1), AAD29441.1; PaPAO1 (Ps. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; VcE1 (V. cholerae E1), P15493.2; PfIFO (P. fragi IFO-12049), P08658.2; PrvK80 (Proteus vulgaris K80), AAB01071.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Ab348935 (A. baumannii 348935), WP_034704719.1; LipG1, this study; Agyl (A. gyllenbergii), WP_032860322.1; Ajun (A. junii), WP_005402474.1. Symbols: ● Catalytic triad residues, ▲ Asp residues involved in calcium binding, ★ HG dipeptide of the oxyanion loop, ○ Putative triad residue, △Putative calcium binding Asp residues.

Mentions: Bioinfomatics analysis showed that the lipG1 gene (KM925083) contained an open reading frame with 1,227 nucleotides, encoding a protein of 406 amino acids (Fig A in S2 File). A 20 amino acid residue signal peptide was found at the N-terminal of the lipase with a cleavage position between Cys20 and His21 as predicted by Signal P (Fig A in S2 File). BLASTp analysis indicated that LipG1 was 99% identical to hypothetical protein from Acinetobacter tandoii (WP_016167430.1), and was 70%, 68%, 68%, 62% and 61% identical to lipases from A. baumannii 348935 (WP_034704719.1), Acinetobacter sp. MII (KGH48978.1), A. lwoffii SH145(WP_004279025), A. gyllenbergii (WP_032860322.1) and A. junii (WP_005402474.1), respectively. Alignment of LipG1 and its relatives with representatives of subfamily I.1 and I.2 of bacterial true lipases revealed the typical lipase semi-conserved pentapeptide, GXSXG, where the catalytic residue Ser is located, as well as the catalytic residue Asp, at homologous positions in all sequences (Fig 1). The last catalytic residue constituting the lipase consensus catalytic triad, His, was not found at homologous position in LipG1. Nevertheless, a conserved His residue was located in LipG1 and its relatives at the position close to the catalytic His for I.1 and I.2 lipases (Fig 1). Therefore, the consensus lipase catalytic triad in LipG1putatively comprises of Ser 190 (in the motif GHSQG), Asp 354 and His 386, which requires further experimental confirmation. An HG sequence, which putatively constitutes an oxyanion hole in the three-dimensional protein structure, was also located at homologous position for all sequences (Fig 1).


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

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

Alignment of amino acid sequences of representative lipases from subfamilies I.1 and I.2 of bacterial lipases and the group of lipases represented by LipG1.Abbreviations and sequence accession numbers: AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; AbBD5 (A. baumannii BD5), ABW70205.1; AcRAG-1 (A. calcoaceticus RAG-1), AAD29441.1; PaPAO1 (Ps. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; VcE1 (V. cholerae E1), P15493.2; PfIFO (P. fragi IFO-12049), P08658.2; PrvK80 (Proteus vulgaris K80), AAB01071.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Ab348935 (A. baumannii 348935), WP_034704719.1; LipG1, this study; Agyl (A. gyllenbergii), WP_032860322.1; Ajun (A. junii), WP_005402474.1. Symbols: ● Catalytic triad residues, ▲ Asp residues involved in calcium binding, ★ HG dipeptide of the oxyanion loop, ○ Putative triad residue, △Putative calcium binding Asp residues.
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Related In: Results  -  Collection

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pone.0132049.g001: Alignment of amino acid sequences of representative lipases from subfamilies I.1 and I.2 of bacterial lipases and the group of lipases represented by LipG1.Abbreviations and sequence accession numbers: AcBD413 (Acinetobacter calcoaceticus BD413), CAA56780.1; AbBD5 (A. baumannii BD5), ABW70205.1; AcRAG-1 (A. calcoaceticus RAG-1), AAD29441.1; PaPAO1 (Ps. aeruginosa PAO1), CAA44997.1; P109 (Pseudomonas sp. 109), P26877.1; VcE1 (V. cholerae E1), P15493.2; PfIFO (P. fragi IFO-12049), P08658.2; PrvK80 (Proteus vulgaris K80), AAB01071.1; Bc3959 (Burkholderia cepacia DSM3959), P22088.2; BgPG1 (Burkholderia glumae PG1), Q05489.1; AspMII (Acinetobacter sp. MII), KGH48978; AlSH145 (A. lwoffii SH145), WP_004279025; Ab348935 (A. baumannii 348935), WP_034704719.1; LipG1, this study; Agyl (A. gyllenbergii), WP_032860322.1; Ajun (A. junii), WP_005402474.1. Symbols: ● Catalytic triad residues, ▲ Asp residues involved in calcium binding, ★ HG dipeptide of the oxyanion loop, ○ Putative triad residue, △Putative calcium binding Asp residues.
Mentions: Bioinfomatics analysis showed that the lipG1 gene (KM925083) contained an open reading frame with 1,227 nucleotides, encoding a protein of 406 amino acids (Fig A in S2 File). A 20 amino acid residue signal peptide was found at the N-terminal of the lipase with a cleavage position between Cys20 and His21 as predicted by Signal P (Fig A in S2 File). BLASTp analysis indicated that LipG1 was 99% identical to hypothetical protein from Acinetobacter tandoii (WP_016167430.1), and was 70%, 68%, 68%, 62% and 61% identical to lipases from A. baumannii 348935 (WP_034704719.1), Acinetobacter sp. MII (KGH48978.1), A. lwoffii SH145(WP_004279025), A. gyllenbergii (WP_032860322.1) and A. junii (WP_005402474.1), respectively. Alignment of LipG1 and its relatives with representatives of subfamily I.1 and I.2 of bacterial true lipases revealed the typical lipase semi-conserved pentapeptide, GXSXG, where the catalytic residue Ser is located, as well as the catalytic residue Asp, at homologous positions in all sequences (Fig 1). The last catalytic residue constituting the lipase consensus catalytic triad, His, was not found at homologous position in LipG1. Nevertheless, a conserved His residue was located in LipG1 and its relatives at the position close to the catalytic His for I.1 and I.2 lipases (Fig 1). Therefore, the consensus lipase catalytic triad in LipG1putatively comprises of Ser 190 (in the motif GHSQG), Asp 354 and His 386, which requires further experimental confirmation. An HG sequence, which putatively constitutes an oxyanion hole in the three-dimensional protein structure, was also located at homologous position for all sequences (Fig 1).

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