Purification and characterization of a sarcoplasmic serine proteinase from threadfin bream Nemipterus virgatus muscle

  • Food Chem. 2019 Jun 30;284:198-204. doi: 10.1016/j.foodchem.2019.01.024.
Jin-Yang Liu  1 ,  Asami Yoshida  2 ,  Yi-Li Gao  3 ,  Kazuya Shirota  4 ,  Yasuhiko Shiina  5 ,  Erika Noguchi  6 ,  Koichi Kuwahara  7 ,  Kiyoshi Osatomi  8
Affiliations
  • 1. Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki 852-8521, Japan. Electronic address: [email protected].
  • 2. Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki 852-8521, Japan. Electronic address: [email protected].
  • 3. Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki 852-8521, Japan. Electronic address: [email protected].
  • 4. Central Research Institute, Maruha Nichiro Corporation, Tsukuba, Ibaraki 300-4295, Japan. Electronic address: [email protected].
  • 5. Central Research Institute, Maruha Nichiro Corporation, Tsukuba, Ibaraki 300-4295, Japan. Electronic address: [email protected].
  • 6. Nagasaki Prefectural Institute of Fisheries, Nagasaki, Taira 851-2213, Japan. Electronic address: [email protected].
  • 7. Nagasaki Prefectural Institute of Fisheries, Nagasaki, Taira 851-2213, Japan. Electronic address: [email protected].
  • 8. Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki 852-8521, Japan. Electronic address: [email protected].
PMID: 30744846 DOI: 10.1016/j.foodchem.2019.01.024
Abstract

A sarcoplasmic serine proteinase (SSP) was purified from threadfin bream (Nemipterus virgatus) belly muscle by ammonium sulfate precipitation and a series of chromatographies including Q-Sepharose, Phenyl Sepharose and Superdex 200. The SSP was purified 1967 folds with a yield of 4.8%. The molecular weight of the SSP was estimated to be 43.5 kDa and 22.5 kDa on SDS-PAGE under non-reducing and reducing conditions, respectively. The N-terminal amino acid sequence of the two protein bands were determined as IVGGYEXQPYSQAHQVSLNSGY and corresponded. It is suggested that the SSP exists as a homodimer. Optimum pH and temperature were 9.5 and 50 °C, using Boc-Val-Pro-Arg-MCA as a substrate. Substrate specificity and effects of inhibitors indicated that the SSP was a trypsin-like serine proteinase. The SSP was responsible for hydrolyzing Myosin heavy chain (MHC) and inducing modori phenomenon in the threadfin bream surimi gel. Thus, the SSP was considered as a modori-inducing proteinase.

Keywords
Modori phenomenon; Purification; Sarcoplasmic fraction; Serine proteinase; Threadfin bream.
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