Abstract
A new enzyme (alkaline protease 894) obtained from the marine extremophile Flavobacterium yellowsea (YS-80-122) has exhibited strong substrate-binding and catalytic activity, even at low temperature, but the characteristics of the hydrolysis with this enzyme are still unclear. The pearl oyster Pinctada martensii was used in this study as the raw material to illustrate the kinetic properties of protease 894. After investigating the intrinsic relationship between the degree of hydrolysis and several factors, including initial reaction pH, temperature, substrate concentration, enzyme concentration, and hydrolysis time, the kinetics model was established. This study showed that the optimal conditions for the enzymatic hydrolysis were an initial reaction pH of 5.0, temperature of 30°C, substrate concentration of 10% (w/v), enzyme concentration of 2 500 U/g, and hydrolysis time of 160 min. The kinetic characteristics of the protease for the hydrolysis of P. martensii were obtained. The inactivation constant was found to be 15.16/min, and the average relative error between the derived kinetics model and the actual measurement was only 3.04%, which indicated a high degree of fitness. Therefore, this study provides a basis for the investigation of the concrete kinetic characteristics of the new protease, which has potential applications in the food industry.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bisswanger H. 2008. Enzyme Kinetics: Principles and Methods (2nd Edn.). The Deutsche Nationalbibliothek. Weinheim. Press, Germany. p.301.
Fawcett J K. 1954. The semi-micro kjeldahl method for the determination of nitrogen. J. Med. Lab. Technol., 12(1): 1–22.
Gonzalez-Tello P, Camacho F, Jurado E, Paez M P, Guadix E M. 1994. Enzymatic hydrolysis of whey proteins: I. kinetic models. Biotechnol. Bioeng., 44(4): 523–528.
Guo X M, Ford S E, Zhang F S. 1999. Molluscan aquaculture in China. J. Shellfish Res., 18(1): 19–31.
Jing G, Li L Y, Li Y, Xie L P, Zhang R Q. 2006. Purification and partial characterization of two acid phosphatase forms from pearl oyster (Pinctada fucata). Comp. Biochem. Phys. B., 143(2): 229–235.
Kanoh S, Aoyama M, Watanabe M, Mae M, Takagi K, Shimomura K, Niwa E. 2001. Possible utilization of the pearl oyster Pinctada Fucata glycogen as a cosmetic material. Nippon. Suisan. Gakk., 67(1): 90–95.
Katano S, Oki T, Matsuo Y, Yoshihira K, Nara Y, Miki T, Matsui T, Matsumoto K. 2003. Antihypertensive effect of alkaline protease hydrolysate of the pearl oyster Pinctada Fucata Martencii & separation and identification of angiotensin-I converting enzyme inhibitory peptides. Nippon. Suisan. Gakk., 69(6): 975–980.
Marquez M C, Vazquez M A. 1999. Modeling of enzymatic protein hydrolysis. Process Biochem., 35(1–2): 111–117.
Mizuta S, Miyagi T, Nishimiya T, Yoshinaka R. 2002. Partial characterization of collagen in mantle and adductor of pearl oyster (Pinctada Fucata). Food Chem., 79(3): 319–325.
Netto F M, Galeazzi M M. 1998. Production and characterization of enzymatic hydrolysate from soy protein isolate. Lebensmittelwissenschaft and Technology, 31(8): 624–631.
Song Y, Mi S, Wang Y, Haiying W, Wei L. 2009. Study on the advantages of marined alkaline protease 894 in the preparation of marine active collagen peptides. Food Sci. Technol., 34(2): 5.
Suetsuna K. 2002. Identification of antihypertensive peptides from peptic digest of the short-necked clam tapes philippinarum and the pearl oyster Pinctada Fucata Martensii. Fisheries Sci., 68(1): 233–235.
Sun M, Xiu Z Y, Wang Y J, Chen L F, Zhang Y B, Hong Y G, Yan X L, Han Y B, Li J, Chen L. 2001. Study on properties of low-temperature protease of Flavobacterium YS-9412-130. Marine Fisheries Research, 22(2): 1–10. (in Chinese with English abstract)
Takakura D, Norizuki M. Ishikawa F, Samata T. 2008. Isolation and characterization of the N-linked oligosaccharides in Nacrein from Pinctada Fucata. Mar. Biotechnol., 10(3): 290–296.
Wang F, Hao J H, Yang C Y, Sun M. 2010. Cloning, expression, and identification of a novel extracellular cold-adapted alkaline protease gene of the marine bacterium strain Ys-80-122. Appl. Biochem. Biotech., 162(5): 1 497–1 505.
Xiao R, Xie L P, Lin J Y, Li C H, Chen Q X, Zhou H M, Zhang R Q. 2002. Purification and enzymatic characterization of alkaline phosphatase from Pinctada fucata. J. Mol. Catal. B-Enzym., 17(2): 65–74.
Xiao R W, Huang L L, Huang K, Cui C, Zhao M M. 2009. Characteristics of Pinctada martensii protein hydrolysates by different proteases. Modern Food Science and Technology, 25(7): 725–730. (in Chinese with English abstract)
Zhang C H, Wu H M, Hong P Z, Deng S G, Lei X L. 2000. Nutrients and composition of free amino acid in edible part of Pinctada martensii. Journal of Fisheries of China, 24: 135–139. (in Chinese with English abstract)
Zhang Z W, Liu X H, Zhou J F, Wu N X, Tong Z F, Liao D K. 2008. Optimation technical conditions for preparing antihypertensive-peptide (ACEI) from Pinctada martensii with alkali protease hydrolysis. Marine Sciences, 32(8): 25–29. (in Chinese with English abstract)
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by the Comprehensive Strategic Cooperation Programs between Guangdong Province and Chinese Academy of Sciences (No. 2011A090100008), and the Knowledge Innovation Program of Chinese Academy of Sciences (No. KZCX2-EW-Q214)
Rights and permissions
About this article
Cite this article
Chen, X., Chen, H., Cai, B. et al. Kinetic study of alkaline protease 894 for the hydrolysis of the pearl oyster Pinctada martensii . Chin. J. Ocean. Limnol. 31, 591–597 (2013). https://doi.org/10.1007/s00343-013-2227-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00343-013-2227-7