Preparation and Antioxidant Activities In Vitro of a Designed Antioxidant Peptide from Pinctada fucata by Recombinant Escherichia coli
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An antioxidant peptide derived from Pinctada fucata meat using an Alcalase2.4L enzymatic hydrolysis method (named AOP) and identified by LC-TOF-MS has promising clinical potential for generating cosmetic products that protect skin from sunshine. To date, there have been few published studies investigating the structure-activity relationship in these peptides. To prepare antioxidant peptides better and improve their stability, the design and expression of an antioxidant peptide from Pinctada fucata (named DSAOP) was studied. The peptide contains a common precursor of an expression vector containing an α-helix tandemly linked according to the BamHI restriction sites. The DNA fragments encoding DSAOP were synthesized and subcloned into the expression vector pET-30a (+), and the peptide was expressed mostly as soluble protein in recombinant Escherichia coli. Meanwhile, the DPPH radical scavenging activity, superoxide radical scavenging activity, and hydroxyl radical scavenging activity of DSAOP IC50 values were 0.136 ± 0.006, 0.625 ± 0.025, and 0.306 ± 0.015 mg/ml, respectively, with 2-fold higher DPPH radical scavenging activity compared with chemosynthesized AOP (p < 0.05), as well as higher superoxide radical scavenging activity compared with natural AOP (p < 0.05). This preparation method was at the international advanced level. Furthermore, pilot-scale production results showed that DSAOP was expressed successfully in fermenter cultures, which indicated that the design strategy and expression methods would be useful for obtaining substantial amounts of stable peptides at low costs. These results showed that DSAOP produced with recombinant Escherichia coli could be useful in cosmetic skin care products, health foods, and pharmaceuticals.
Abstract
An antioxidant peptide derived from Pinctada fucata meat using an Alcalase2.4L enzymatic hydrolysis method (named AOP) and identified by LC-TOF-MS has promising clinical potential for generating cosmetic products that protect skin from sunshine. To date, there have been few published studies investigating the structure-activity relationship in these peptides. To prepare antioxidant peptides better and improve their stability, the design and expression of an antioxidant peptide from Pinctada fucata (named DSAOP) was studied. The peptide contains a common precursor of an expression vector containing an α-helix tandemly linked according to the BamHI restriction sites. The DNA fragments encoding DSAOP were synthesized and subcloned into the expression vector pET-30a (+), and the peptide was expressed mostly as soluble protein in recombinant Escherichia coli. Meanwhile, the DPPH radical scavenging activity, superoxide radical scavenging activity, and hydroxyl radical scavenging activity of DSAOP IC50 values were 0.136 ± 0.006, 0.625 ± 0.025, and 0.306 ± 0.015 mg/ml, respectively, with 2-fold higher DPPH radical scavenging activity compared with chemosynthesized AOP (p < 0.05), as well as higher superoxide radical scavenging activity compared with natural AOP (p < 0.05). This preparation method was at the international advanced level. Furthermore, pilot-scale production results showed that DSAOP was expressed successfully in fermenter cultures, which indicated that the design strategy and expression methods would be useful for obtaining substantial amounts of stable peptides at low costs. These results showed that DSAOP produced with recombinant Escherichia coli could be useful in cosmetic skin care products, health foods, and pharmaceuticals.
참고문헌 (42)
- Ahn C, "Purification and antioxidant properties of octapeptide from salmon byproduct protein hydrolysate by gastrointestinal digestion", Food Chem, 147, pp.78~83 (2014)[DOI]
- Zou T, "The structureactivity relationship of the antioxidant peptides from natural proteins", Molecules, 21, pp.72~ (2016)[DOI]
- Zambrowicz A, "Multifunctional peptides derived from an egg yolk protein hydrolysate: isolation and characterization", Amino Acids, 47, pp.369~380 (2015)[DOI]
- Zhuang H, "Purification and identification of antioxidant peptides from corn gluten meal", J. Funct. Foods, 5, pp.1810~1821 (2013)
- Chen C, "Purification and identification of antioxidant peptides from egg white protein hydrolysate", Amino Acids, 43, pp.457~466 (2012)[DOI]
- Liu Q, "Antioxidant activity and functional properties of porcine plasma protein hydrolysate as influenced by the degree of hydrolysis", Food Chem, 118, pp.403~410 (2010)
- Guo H, "Structures and properties of antioxidative peptides derived from royal jelly protein", Food Chem, 113, pp.238~245 (2009)[DOI]
- Cheung IW, "The role of molecular size in antioxidant activity of peptide fractions from Pacific hake (Merluccius productus) hydrolysates", Food Chem, 134, pp.1297~1306 (2012)[DOI]
- Nalinanon S, "Functionalities and antioxidant properties of protein hydrolysates from the muscle of ornate threadfin bream treated with pepsin from skipjack tuna", Food Chem, 124, pp.1354~1362 (2011)
- Zhong S, "Antioxidant properties of peptide fractions from silver carp (Hypophthalmichthys molitrix) processing by-product protein hydrolysates evaluated by electron spin resonance spectrometry", Food Chem, 126, pp.1636~1642 (2011)[DOI]
- Wu YY, "Application of response surface methodology to optimise preparation high antioxidant activity product from Pinctada fucata muscle.", Adv. Mater. Res., 396-398, pp.1341~1348 (2012)
- Wu YY, "Inhibitory effect of antioxidant peptides derived from Pinctada fucata protein on ultraviolet-induced photoaging in mice", J. Funct. Foods, 5, pp.527~538 (2013)
- Wu YY, "Desalination of antioxidant peptides from Pinctada fucata using C18 chromatographic column", Adv. Mater. Res., 781-784, pp.1508~1512 (2013)[DOI]
- Farvin KHS, "Antioxidant activity of yoghurt peptides: part 2 - characterisation of peptide fractions", Food Chem, 123, pp.1090~1097 (2010)[DOI]
- Axarli I, "Directed evolution of Tau class glutathione transferases reveals a site that regulates catalytic efficiency and masks co-operativity", Biochem. J., 473, pp.559~570 (2016)[DOI]
- Tong Y, "Novel integration strategy coupling codon and fermentation optimization for efficiently enhancing sarcosine oxidase (SOX) production in recombinant Escherichia coli", World J. Microbiol. Biotechnol., 31, pp.707~716 (2015)[DOI]
- Memarpoor-Yazdi M, "Antioxidant activity of protein hydrolysates and purified peptides from Zizyphus jujuba fruits", J. Funct. Foods, 5, pp.62~70 (2013)[DOI]
- Xia Y, "Fractionation and characterization of antioxidant peptides derived from barley glutelin by enzymatic hydrolysis", Food Chem, 134, pp.1509~1518 (2012)[DOI]
- Zhuang H, "Optimisation of antioxidant peptide preparation from corn gluten meal", J. Sci. Food Agric., 93, pp.3264~3270 (2013)[DOI]
- Anane E, "Optimisation and scale-up of <TEX>${\alpha}$<", TEX>-glucuronidase production by recombinant Saccharomyces cerevisiae in aerobic fed-batch culture with constant growth rate, 2013, pp.81, 1~7
- Duan X, "Purification and characterization of a natural antioxidant peptide from fertilized eggs", Food Res. Int., 56, pp.18~24 (2014)[DOI]
- Yang F, "Expression and purification of Canis interferon <TEX>${\alpha}$<", TEX> in Escherichia coli using different tags, 2015, pp.115, 76~82
- Bis RL, "High yield soluble bacterial expression and streamlined purification of recombinant human interferon <TEX>${\alpha}$<", TEX>-2a, 2014, pp.99, 138~146
- Kralicek A., "A cell-free expression screen to identify fusion tags for improved protein expression", Methods Mol. Biol., 1118, pp.35~ (2014)
- Parachin NS, "Expression systems for heterologous production of antimicrobial peptides", Peptides, 38, pp.446~456 (2012)
- Vargas-Cortez T, "Expression and purification of recombinant proteins in Escherichia coli tagged with a small metal-binding protein from Nitrosomonas europaea", Protein Expr. Purif., 118, pp.49~54 (2016)[DOI]
- Wei Hu, "Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensis", Journal of Microbiology and Biotechnology, 27(5), pp.965~974 (2017), 한국미생물·생명공학회[DOI]
- Zou W, "Expression, purification, and characterization of recombinant human L-chain ferritin", Protein Expr. Purif., 119, pp.63~68 (2016)[DOI]
- Wang Q, "Purification and characterization of aspartate N-acetyltransferase: a critical enzyme in brain metabolism", Protein Expr. Purif., 119, pp.11~18 (2016)[DOI]
- Jin D, "Preparation of antioxidative corn protein hydrolysates, purification and evaluation of three novel corn antioxidant peptides", Food Chem, 204, pp.427~436 (2016)[DOI]
- Slizyte R, "Functional, bioactive and antioxidative properties of hydrolysates obtained from cod (Gadus morhua) backbones", Process Biochem., 44, pp.668~677 (2009)[DOI]
- Jia J, "Enzymatic hydrolysis of Alaska pollack (Theragra chalcogramma) skin and antioxidant activity of the resulting hydrolysate", J. Sci. Food Agric., 90, pp.635~640 (2010)
- Hathwar SC, "Simultaneous recovery of lipids and proteins by enzymatic hydrolysis of fish industry waste using different commercial proteases", Appl. Biochem. Biotechnol., 164, pp.115~124 (2011)
- Chi C, "Isolation and characterization of three antioxidant peptides from protein hydrolysate of bluefin leatherjacket (Navodon septentrionalis) heads", J. Funct. Foods, 12, pp.1~10 (2015)[DOI]
- Cheung IW, "The role of molecular size in antioxidant activity of peptide fractions from Pacific hake (Merluccius productus) hydrolysates", Food Chem., 134, pp.1297~1306 (2012)[DOI]
- Li HM, "Antioxidant properties and possible mode of action of corn protein peptides and zein peptides", J. Food Biochem., 34, pp.44~60 (2010)[DOI]
- Je J, "Antioxidant and antihypertensive protein hydrolysates produced from tuna liver by enzymatic hydrolysis", Food Res. Int., 42, pp.1266~1272 (2009)
- Ngo D, "In vitro antioxidant activity of a peptide isolated from Nile tilapia (Oreochromis niloticus) scale gelatin in free radical-mediated oxidative systems", J. Funct. Foods, 2, pp.107~117 (2010)[DOI]
- Samaranayaka AG, "Food-derived peptidic antioxidants: a review of their production, assessment, and potential applications", J. Funct. Foods, 3, pp.229~254 (2011)[DOI]
- Sharma OP, "DPPH antioxidant assay revisited", Food Chem, 113, pp.1202~1205 (2009)[DOI]
- Jhamb K, "Production of soluble recombinant proteins in Escherichia coli: effects of process conditions and chaperone co-expression on cell growth and production of xylanase", Bioresour. Technol., 123, pp.135~143 (2012)[DOI]
- Pal G, "Scaling up the production of recombinant antimicrobial plantaricin E from a heterologous host, Escherichia coli", Probiotics Antimicrob. Proteins, 7, pp.216~221 (2015)[DOI]