title: Matrix Metalloproteinase 2 (MMP-2) Plays a Critical Role in the Softening of Common Carp Muscle during Chilled Storage by Degradation of Type I and V Collagens.
abstract: Matrix metalloproteinases (MMPs) are proposed to play important roles in the degradation of collagens, thus causing the post-mortem softening of fish muscle, although the specific mechanism remains largely unresolved. Previously, we reported the existence of gelatinase-like proteinases in common carp (Cyprinus carpio) muscle. The primary structures of these proteinases, however, have never been investigated. In the present study, two MMPs with molecular masses of 66 and 65 kDa were purified to homogeneity from common carp muscle by ammonium sulfate fractionation and a series of column chromatographies. Matrix-assisted laser desorption/ionization-tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS/MS) analysis indicated that they are completely identical to MMP-2 from common carp. During chilled storage of common carp at 4 °C, the enzymatic activity of MMP-2 increased to 212% in 12 h while the texture profile increased over the first 2 h and gradually decreased. On the other hand, type V collagen was purified to homogeneity and a specific polyclonal antibody against this protein was prepared. Both type I and V collagens were effectively hydrolyzed by MMP-2 at 30 °C and even at 4 °C. Furthermore, injection of metalloproteinase proteinase inhibitor EDTA into the blood vessel of live common carp suppressed post-mortem tenderization significantly. All of these results confirmed that MMP-2 is a major proteinase responsible for the degradation of collagens, resulting in the softening of fish muscle during chilled storage.
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.
authors.forename: Chao
authors.initials: C
authors.lastname: Xu
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.
authors.forename: Cheng
authors.initials: C
authors.lastname: Wang
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Qiu-Feng
authors.initials: QF
authors.lastname: Cai
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Ling
authors.initials: L
authors.lastname: Weng
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Guang-Ming
authors.initials: GM
authors.lastname: Liu
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Wen-Jin
authors.initials: WJ
authors.lastname: Su
authors.affiliation: College of Food and Biological Engineering, Jimei University , Xiamen, China 361021.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources , Xiamen, Fujian, China 361102.
authors.forename: Min-Jie
authors.initials: MJ
authors.lastname: Cao
mesh_terms: D000818:Animals; D002347:Carps; D003080:Cold Temperature; D003094:Collagen / Q000378:metabolism*; D024042:Collagen Type I / Q000378:metabolism; D024062:Collagen Type V / Q000378:metabolism; D005519:Food Preservation / Q000379:methods*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology / Q000378:metabolism*; D017747:Seafood / Q000032:analysis*; D012677:Sensation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D024042:Collagen Type I; D024062:Collagen Type V; D003094:Collagen; D020778:Matrix Metalloproteinase 2
keywords: gelatin zymography; matrix metalloproteinase-2 (MMP-2); purification; texture profiles; type V collagen