h 0 ~ 4 Table 1 Design of different treatment groups Alcalase2. 4 L > U /g 11 ~ 250 kda Promega 5. 5 g 0. 5 g Foss 1. 2 Allegra

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32 7 2 0 1 6 7 Vol. 32 No. 7 Jul. 2 0 1 6 DOI 10. 13652 /j. issn. 1003-5788. 2016. 07.

1 Vol. 32 No. 7 Jul DOI /j. issn Curdlan Alcalase Optimization on processing of curdlan and alcalase to chilled prepared beef LONG Men MEN Jia-li ZHAN Ge DU Qing-fei ZHOU Di CAI Hua-zhen School of Bio and Food Engineering Chuzhou University Chuzhou Anhui China Curdlan Alcalase ring time h from response surface results by quadratic regression and best tender process was Curdlan mg /g Alcalase U /g and cu- through the result of verification test and the shear force cooking loss Curdlan sensory evaluation was ± kg ± % ± respectively. The relative error is less than 5% which compared with the theoretical value. Then the processes were accurate and P < apply to chilled prepared beef. Alcalase Keywords beef Curdlan Alcalase fresh food tenderization proteolysis optimization Curdlan Alcalase P < Curdlan Alcalase 1 Curdlan mg /g Alcalase U /g h ± kg ± Nisin 5% % ± Curdlan β-l 3-D- Abstract Fresh beef was used to study the effects of Curdlan and alcalase 2006 on shear force cooking loss and sensory quality of chilled prepared beef 6 ph then the process was optimized. The result showed that compared the 3. 0 ~ 9. 5 control Curdlan could has significant effect on the shear force cooking 7 - Alcalase loss and sensory quality of chilled prepared beef P < but no significant Ala on protein and alcalase had a significant hydrolysis of myofibrillar 9 - Leu Val Tyr Phe Trp protein. Different level of Curdlan and alcalase had a significant P < Curdlan 05 impact on the shear force cooking loss of chilled prepared beef and Alcalase could improve the sensory quality significantly P < At last the Curdlan Alcalase 2014GH44 KJ2016SD jxtd zhangeczxy@ 163. com GB /T

2 h 0 ~ 4 Table 1 Design of different treatment groups Alcalase2. 4 L > U /g 11 ~ 250 kda Promega 5. 5 g 0. 5 g Foss 1. 2 Allegra 64R Beckman Coulter Kjeltec TM 2300 KDN-04 Foss Bio-Rad 2 Curdlan Alcalase ZKSY U /g mg /g Curdlan 4 72 h AUY120 Shimadzu UV Alcalase Curdlan ng /g U /g Alcalase h T 1 Alcalase Curdlan 4 4 Design Expert T 2 Alcalase 4 State-Ease Minneapolis MN USA Box-Behnken T 3 Curdlan 4 Curdlan Alcalase T cm 3 2 SDS-PAGE 16 cm 1 cm 3 17 Alcalase % Alcalase Microsoft Excel SAS 9. 2 Curdlan Curdlan 10% ANOVA LSD 50 3 min Design Expert min % % 0. 01% % % 0. 01% 172 Curdlan / Alcalase / mg g - 1 U g - 1 T T T T Foss 1 Alcalase 4 U /g Curdlan 5 mg /g h Alcalase Curdlan Curdlan Alcalase 2. 1 Curdlan Alcalase 1 Curdlan Alcalase T 4 Curdlan Alcalase 4. 0 mg /g 3 U /g Curdlan T 3 P < T 1 T 2 T 3 T h Alcalase P < T 2 Curdlan Alcalase T 1 2 Curdlan Alcalase T 2 P > Curdlan Alcalase 1 Alcalase 4 48 h SDS-PAGE P < T 3 Curdlan Curdlan Alcalase

32 7 Curdlan Alcalase P < 0. 05 1 72 h Figure 1 Effect of different salting conditions on chilled prepared beef 2. 2. 2 Curdlan 4 P < 0. 05 Curdlan Alcalase P < 0.

3 32 7 Curdlan Alcalase P < h Figure 1 Effect of different salting conditions on chilled prepared beef Curdlan 4 P < Curdlan Alcalase P < Curdlan Curdlan Curdlan Alcalase Curdlan T 1 3 P < T 4 Alcalase 120 ~ 200 kda 70 kda 40 ~ 50 kda T 1 T 2 T 3 Curdlan T 1 T 2 85 ~ 200 kda 30 ~ 40 kda 40 ~ 50 kda T 3 Curdlan 9-10 Curdlan Alcalase ~ 72 h P < Curdlan Alcalase Alcalase Curdlan 1 Curdlan 3 Figure 3 Effect of different salting time on 8 chilled prepared beef 2 Alcalase 2 SDS-PAGE Figure 2 SDS-PAGE images of protein of chilled prepared beef 4 Curdlan Figure 4 Effect of different Curdlan dosage on chilled prepared beef 173

4 Curdlan Curdlan Table 3 The result of response surface experiment X 1 X 2 X 3 /kg /% Alcalase Alcalase U /g P < Alcalase Alcalase 4 Figure 5 Effect of different Alcalase dosage on Table 4 Test for significance of regression coefficients chilled prepared beef F P F P F P < < Design-Expert X < Y 1 X Y 2 Y 3 3 X Y 1 = X X X X 1 X 2 + X 1 X 2 X 1 X X 1 X X 2 X X X X 2 X X X < < Y 2 = X X X X 1 X X 1 X X 2 X X X X Y 3 = X X X X 1 X X 1 X X 2 X X X X F R 2 Adj P <0. 01 P > R 2 4 Alcalase Curdlan 2 Table 2 Codes and levels of factors chosen for experiment Alcalase Curdlan X 1 Curdlan X 2 Alcalase X 3 / Curdlan / mg g - 1 / U g - 1 h Alcalase a X < X < R Adj-R

5 32 7 Curdlan Alcalase 6 7 Figure 6 Effect of different factors interaction on the shear of chilled prepared beef Figure 7 Effect of different factors interaction on the cooking loss of chilled prepared beef 72 h Alcalase Curdlan Curdlan Alcalase 6 b 70 ~ 80 h Alcalase 2 7 Curdlan 5. 5 mg /g Alcalase 72 h Alcalase Curdlan Curdlan Curdlan 8 Figure 8 Effect of different factors interaction on the 3 8 sensory of chilled prepared beef 175

6 Curdlan protein with curdlan and transglutaminase J. Food Chemistry Curdlan h Alcalase 8. J Curdlan 9 QUAGLIA G B ORBAN E. Influence of the degree of hydrolysis on the solubility of the protein hydrolysates from sardine J. J. Sci. Food Agricultral Alcalase 4 U /g Curdlan 10 GUERARD F DUFOSSE L BROISE D D L et al. Enzymatic hydrolysis of proteins from yellowfin tuna Thunnus albacares wastes using Alcalase J. J. Mol. Catal B Enzym Design-Expert 11 FONKWE L G SINGH R K. Protein recovery from mechanically deboned turkey residue by enzymic hydrolysis J. Process Biochemical Curdlan mg /g Alcalase U /g h 12. Alcalase 1. 0 kg % 92. J ± kg 13. Alcalase ± % ± n = 3 J % 3 14 KOTZEKIDOU P BLOUKAS J G. Microbial and sensory changes in vacuum-packed frankfurter-type sausage by Lactobacillus alimentarius and fate of inoculated Salmonella enteritidis J. Food Microbiology Curdlan Alcalase Alcalase 15. Curdlan J Curdlan Alcalase P 16 SUN Yang-ying PAN Dao-dong GUO Yu-xing et al. Purification < Curdlan Alcalase of chicken breast protein hydrolysate and analysis of its antioxidant activity J. Food and Chemical Toxicology J. procedure J. Food Quality & Preference CAINE W R AALHUS J L BEST D R et al. Relationship of texture profile analysis and Warner-Bratzler shear force with sensory D characteristics of beef rib steaks J. Meat Science J ELMASRY G SUN D W ALLEN P. Chemical-free assessment and 19 FUNAMI T UNAMI M YADA H et al. Gelation mechanism of mapping of major constituents in beef using hyperspectral imaging curdlan by dynamic viscoelasticity measurements J. Journal of J. Journal of Food Engineering ZHAN Xiao-bei LIN Chi-chung ZhANG Hong-tao. Recent advances in curdlan biosynthesis biotechnological production and applications J. Applied Microbiology and Biotechnology YAN Jing-kun MA Hai-le CAI Pan-fu et al. Structural characteristics and antioxidant activities of different families of 4-acetamido- TEMPO-oxidised curdlan J. Food Chemistry HU Ya-qin LIU Wen-juan YUAN Chun-hong et al. Enhancement of the gelation properties of hairtail Trichiurus haumela muscle AASLYNG M D BEJERHOLM C ERTBIERG P et al. Cooking loss and juiciness of pork in relation to raw meat quality and cooking Food Science

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g

H 2 SO ml ml 1. 0 ml C 4. 0 ml - 30 min 490 nm 0 ~ 100 μg /ml Zhao = VρN 100% 1 m V ml ρ g 16 6 2018 11 Chinese Journal of Bioprocess Engineering Vol. 16 No. 6 Nov. 2018 doi 10. 3969 /j. issn. 1672-3678. 2018. 06. 004 1 2 1 1 1. 330200 2. 330006 4 Box-Behnken 1 25 g /ml 121 92 min 2 6. 284%