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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

Characterization of purified recombinant LipG1.a. The effect of pH on lipase activity. The activity assay was performed at 37°C in buffers of pH 3.0–12.0 for 10 min. b. The effect of temperature on lipase activity measured in 0.1 M PBS buffer (pH 8.0) for 10 min. c. pH stability of LipG1. After pre-incubating the enzyme at 37°C for 1 h at various pHs, the residual activity was measured in 0.1 M PBS buffer (pH 8.0), 40°C. d. Thermostability of purified LipG1. The enzyme was preincubated at 20, 30, 40, or 50°C in 0.1 M PBS buffer (pH 8.0). Aliquots were removed at specific time points for measurement of residual activity in the same buffer at 40°C. e. Substrate specificity of LipG1 determined with various chain length fatty acid esters. f. Effects of proteases on the activity of LipG1. Each value in the panel represents the mean ± SD (n = 3).
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pone.0132049.g002: Characterization of purified recombinant LipG1.a. The effect of pH on lipase activity. The activity assay was performed at 37°C in buffers of pH 3.0–12.0 for 10 min. b. The effect of temperature on lipase activity measured in 0.1 M PBS buffer (pH 8.0) for 10 min. c. pH stability of LipG1. After pre-incubating the enzyme at 37°C for 1 h at various pHs, the residual activity was measured in 0.1 M PBS buffer (pH 8.0), 40°C. d. Thermostability of purified LipG1. The enzyme was preincubated at 20, 30, 40, or 50°C in 0.1 M PBS buffer (pH 8.0). Aliquots were removed at specific time points for measurement of residual activity in the same buffer at 40°C. e. Substrate specificity of LipG1 determined with various chain length fatty acid esters. f. Effects of proteases on the activity of LipG1. Each value in the panel represents the mean ± SD (n = 3).

Mentions: LipG1showed maximum activity at pH 8.0, and retained at least 70% of its maximum activity between pH 7.0 and 8.5 (Fig 2a). LipG1 was stable over a wide pH range, remaining at least 60% of its maximum activity after incubation at pH from 3.0 to 12.0 (Fig 2c). The enzyme activity of LipG1 peaked at 40°C, and retained over 40% of its maximum activity at temperatures from 20°C to 35°C (Fig 2b). The recombinant lipase showed high stability at 20°C and 30°C. However, the activity decreased quickly when incubated at 50°C (Fig 2d).


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

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

Characterization of purified recombinant LipG1.a. The effect of pH on lipase activity. The activity assay was performed at 37°C in buffers of pH 3.0–12.0 for 10 min. b. The effect of temperature on lipase activity measured in 0.1 M PBS buffer (pH 8.0) for 10 min. c. pH stability of LipG1. After pre-incubating the enzyme at 37°C for 1 h at various pHs, the residual activity was measured in 0.1 M PBS buffer (pH 8.0), 40°C. d. Thermostability of purified LipG1. The enzyme was preincubated at 20, 30, 40, or 50°C in 0.1 M PBS buffer (pH 8.0). Aliquots were removed at specific time points for measurement of residual activity in the same buffer at 40°C. e. Substrate specificity of LipG1 determined with various chain length fatty acid esters. f. Effects of proteases on the activity of LipG1. Each value in the panel represents the mean ± SD (n = 3).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4492967&req=5

pone.0132049.g002: Characterization of purified recombinant LipG1.a. The effect of pH on lipase activity. The activity assay was performed at 37°C in buffers of pH 3.0–12.0 for 10 min. b. The effect of temperature on lipase activity measured in 0.1 M PBS buffer (pH 8.0) for 10 min. c. pH stability of LipG1. After pre-incubating the enzyme at 37°C for 1 h at various pHs, the residual activity was measured in 0.1 M PBS buffer (pH 8.0), 40°C. d. Thermostability of purified LipG1. The enzyme was preincubated at 20, 30, 40, or 50°C in 0.1 M PBS buffer (pH 8.0). Aliquots were removed at specific time points for measurement of residual activity in the same buffer at 40°C. e. Substrate specificity of LipG1 determined with various chain length fatty acid esters. f. Effects of proteases on the activity of LipG1. Each value in the panel represents the mean ± SD (n = 3).
Mentions: LipG1showed maximum activity at pH 8.0, and retained at least 70% of its maximum activity between pH 7.0 and 8.5 (Fig 2a). LipG1 was stable over a wide pH range, remaining at least 60% of its maximum activity after incubation at pH from 3.0 to 12.0 (Fig 2c). The enzyme activity of LipG1 peaked at 40°C, and retained over 40% of its maximum activity at temperatures from 20°C to 35°C (Fig 2b). The recombinant lipase showed high stability at 20°C and 30°C. However, the activity decreased quickly when incubated at 50°C (Fig 2d).

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