한수지 51(4), $, 218 Original Article Korean J Fish Aquat Sci 51(4),$,218 해조류첨가가쌀코지제조와효소활성에미치는영향 전준영 이미향 1 정인학 1 정민정 김병목 한국식품연구원, 1 강릉원주대학교해양식품공학과 Effects of Seaweeds on Rice Koji Production and Enzyme Activity Joon-Young Jun, Mi-Hyang Lee 1, In-Hak Jeong 1, Min-Jeong Jung and Byoung-Mok Kim Division of Strategic Food Research, Korea Food Research Institute, Gangneung 2544, Korea 1 Department of Marine Food Science and Technology, Gangneung-Wonju National University, Gangneung 25457, Korea This study investigated the effects of selected seaweeds on rice koji preparation (rice inoculated with Aspergillus luchuensis) and enzyme activity. Four types of rice koji were prepared using one of three seaweeds (.5% laver,.5% kelp and.5% green laver) or a control for 72 h. The changes in the moisture content, water activity, ph, total mold cell count, amylase and protease activities were measured. During preparation, there was no significant difference in the moisture content among the four kojis, whereas the ph in the kojis made with either laver or green laver decreased rapidly compared with the control (P<.5). This seemed to result from the seaweeds promoting the growth of mold cells. In terms of the activities of both amylase and protease, the koji with laver was superior. Subsequently, the amylase and protease activities of the koji made with laver were evaluated at various phs (3 to 9), temperatures (15-55 ), and NaCl concentrations (-1%). The activities of both enzymes decreased notably at ph 9 and the protease activity decreased at temperatures above 45. Although the activities of both enzymes decreased at greater than 2.5% NaCl, activity was present at 1% NaCl. Key words: Rice koji, Aspergillus luchuensis, Seaweed, Enzyme activity, NaCl 서론,,, (Mok et al., 25).,,,,,, (Kim and Lyu, 21).,, (Lim, 28; Park et al., 27). (koji), ( ).,, (Kim et al., 212). Aspergillus luchuensis (syn. A. kawachii) Aspergillus niger amylase, maltase, A. oryzae ph (Kim et al., 213). amylase protease (So and Lee, 21). (Kwon et al., 213), (Lee et al., 211), (Song and Lee, 213), (Kim et al., 215b), seed mash (Lee et al., 25), Aspergillus (Choi et al., 211), soybean koji rice koji (Jun et al., 216), https://doi.org/1.5657/kfas.218. Korean J Fish Aquat Sci 51(4), August 218 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3./) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 14 May 218; Revised 7 June 218; Accepted 19 June 218 Corresponding author: Tel: +82. 33. 643. 842 Fax: +82. 33. 643. 839 E-mail address: bmkim@kfri.re.kr Copyright 218 The Korean Society of Fisheries and Aquatic Science 1 pissn:374-8111, eissn:2287-8815
2 전준영ㆍ이미향ㆍ정인학ㆍ정민정ㆍ김병목 Aspergillus kawachii (Kim et al., 215a),. ( ),.,, amylase protease, ph,, NaCl amylase protease.. 재료및방법 재료및종균 216. Aspergillus luchuensis (Ulsan, Republic of Korea)., 4 mesh.. 쌀코지제조 Jun et al. (216)., (1 kg) 16, 4 3.5% (w/w).. 15 4, UV,.2% A. luchuensis 3. (85%) (3 ) 72. 시료의수집및조효소액추출 72 12 1 g,,, ph.. 2 g 1 ml 1, 3 3. (1, g, 4, 1 ). 수분함량및수분활성도 (925.45; AOAC, 25)., 5 g 2 1, (HP23-AW-A, Rotronic, Switzerland). ph 및곰팡이수 ph, 5 g 45 ml 1, (7, g, 15 ) ph meter (CH/Seven Easy-s2K, Mettler Toledo, Switzerland). (Park and Oh, 1995). 2 g.1% peptone water 18 ml (26 rpm, 5 ),. 1 ml potato dextrose agar (Difco; Becton Dckinson, Spark, MD, USA), 28 72 colony (15-15) log CFU/g. Amylase 활성 amylase Bechman et al. (212) 3,5-dinitrosalicylic acid..1 M acetate buffer (ph 5.) 1% soluble starch 1 ml 1 ml 55 15..5 ml, 96 mm 3,5-dinitrosalicylic acid.5 ml 95 5. 4 ml 15, 54 nm. maltose.2% maltose, standard curve. Amylase 1 g 1 mg maltose 1 unit (U). Protease 활성 Protease O-phthalaldehyde (Shakerian et al., 214)..21 M sodium phosphate buffer (ph 8.2).5% casein 1 ml 1 ml 35 3..75% trichloroacetic acid 4 ml, (7, g, 1 ).
해조류첨가쌀코지의효소활성 3 2 µl O-phthalaldehyde reagent 3 ml 3 1 3 34 nm..5% serine standard curve. Protease 1 g serine 1 µg 1 U. O-phthalaldehyde reagent O-phthaladehyde 8 mg methanol 2% SDS 5 ml, 1 mm sodium tetraborate decahydrate 5 ml, 2-mercaptoethanol 3 µl 1 ml.. ph, 온도및 NaCl 농도가 amylase 와 protease 활성에미치는영향, ph, NaCl. ph 3, 5, 7, 9, 15, 25, 35, 45, 55, NaCl, 2.5, 5, 7.5, 1%. ph. ph 3,.1 M glycine-hcl buffer; ph 5,.1 M acetate buffer; ph 7,.1 M potassium phosphate buffer; ph 9,.1 M tris-hcl buffer. NaCl NaCl. ph (1 ) NaCl (%), NaCl (1 ) (35 ). amylase protease. 통계분석, IBM SPSS statistics program (Version 23; IBM Co., Amork, NY, USA) one-way ANOVA 95% Tukey s test. 결과및고찰 수분함량및수분활성도,.1-1% (72 ) amylase protease,,.5% (data not shown)..5%, Table 1.,,., 72 (P<.5). Aspergillus.65 (Stevenson et al., 215), 72 22.1-27.1%,.78-.84. ph 및곰팡이수 shochu A. kawachii ph ( ph 3.), citric acid (Mikami et al., 1987; Nagamine et al., 23). Kim et al. (213) A. oryzae citric acid, malic acid, lactic acid (dominant), A. kawachii citric acid, succinic acid Table 1. Changes in the moisture content and water activity of the rice kojis with different seaweeds during solid-state fermentation Incubation time (h) Control Laver Kelp Green laver Moisture (%) Water activity Moisture (%) Water activity Moisture (%) Water activity Moisture (%) Water activity 43.6±6.9 A ns.98±.1 A ns 49.5±3.6 A.98±.1 A 43.2±9.5 A.96±.1 A 39.1±3.6 A.99±.1 A 12 4.3±7.5 AB ns.95±.3 AB ns 46.4±6.6 AB.98±.1 A 42.±2.6 A.97±.1 A 32.4±5.8 AB.99±.1 A 24 31.3±4.3 ABC ns.89±.2 ABC a 4.±6. ABC.91±.2 B a 32.3±2.8 AB.9±.2 AB a 31.4±6.2 AB.84±.1 B b 36 29.3±1.1 ABC ns.88±.5 ABC ns 36.±9.6 ABC.87±.3 BC 24.7±9.5 B.9±.5 AB 28.1±3.4 AB.83±.1 B 48 27.4±6.8 BC ns.85±.8 BC ns 31.7±4.5 BC.84±.1 CD 24.8±6.7 B.88±.3 B 28.3±6. AB.79±.2 C 6 26.±1.3 BC ns.81±.1 C bc 29.1±3.2 C.83±.1 CD ab 23.7±2.6 B.86±.2 B a 26.3±3.7 AB.79±.1 C c 72 25.7±3.6 C ns.81±.1 C ab 27.1±5.5 C.82±.2 D ab 22.1±2.1 B.84±.3 B a 24.2±5.5 B.78±.1 C b The different capital letters indicate significantly different values in the vertical column (P<.5). No significant difference in the horizontal column (P<.5). The different small letters indicate significantly different values in the horizontal column (P<.5). Data expressed as the mean±sd. in triplicate.
4 전준영ㆍ이미향ㆍ정인학ㆍ정민정ㆍ김병목 8 1 ph 7 6 5 4 3 Mold cell count (log CFU/g koji) 9 8 7 6 5 2 12 24 36 48 6 72 4 12 24 36 48 6 72 Fig. 1. Changes in the ph and mold cell count of the rice kojis with different seaweeds during solid-state fermentation. Symbols indicate:, control (without seaweed);, laver;, kelp;, green laver. Significant difference at P<.5 versus the control (ANOVA). Data expressed as the mean±sd. in triplicate. oxalic acid., ph. 225 ph Fig. 1. ph 2 (ph 6.6-6.7), 175 36 ph (2.8 )., 12 15 24 125 (P<.5) ph..2% 1 (w/w) 6 log CFU/g (wet basis), 24 75 5. 36 25, 7.6 log CFU/ g 8.2 log 12 CFU/g 24 36 (P<.5). 48 6 72 ph 12 Incubation time (h),, life cycle.,,, (Madigan and Martinko, 26), ph 2. Amylase activity (U/g koji) Amylase 및 protease 활성 Amylase activity (U/g koji) 175 15, 125 1 75 5,,, 12. Bechman et al. (212), 1 A. oryzae 58 -amylase. 8 72, amylase 72 6 (Fig. 2A). Amylase 36, 4 (P<.5)., 36 72 2 (P<.5), amylase 1 g 1,74 U, 1,733 U, 1,586 U, 1,378 U 12. 24 36 48 6 72 Amylase 72 Incubation time (h), protease 36 (Fig. 2B). 24, 36 (P<.5). 7% 9% (Choi, 2 21), protease,a a,b ab,bc ab,c b,c b amylase, A a, B a b 175, A b, B b, B b, B b. Aspergillus 15, Protease activity (U/g koji) Amylase activity (U/g koji) 125 1 75 5
해조류첨가쌀코지의효소활성 5 225 12 Amylase Amylase activity activity (U/g koji) (U/g koji) 2 225 175 2 15 175 125 15 1 125 1 75 5 75 25 5 25 12 24 36 48 6 72 12 24 Incubation 36 time (h) 48 6 72 Protease Protease activity activity (U/g koji) (U/g koji) 12 1 1 8 8 6 6 4 4 2 2 12 24 36 48 6 72 12 24 Incubation 36time (h) 48 6 72 Fig. 2. Changes in the amylase and protease activities of the rice kojis with different seaweeds during solid-state fermentation. Symbols indicate:, control (without seaweed);, laver;, kelp;, green laver. One unit of amylase activity was defined as the one milligram of maltose released from 1% soluble starch per a minute. One unit of protease activity was defined as the one microgram of leucine released from.5% casein per a minute. Significant difference at P<.5 versus the control (ANOVA). Data expressed as the mean±sd. in triplicate. Amylase Amylase activity activity (U/g koji) (U/g koji) 2 175 2 15 175 125 15 1 125 1 75 5 75 25 5 25,C b,b a,a a,a a,a a,,c B ab,,b B a,,a B ab,,a A a,,a A a, b,, B AB a b,, A AB a c,, A AB a b, b, B b 15, B b 25, AB b 35, AB c 45, AB b 55, A b 15 25 Temperature 35 ( ) 45 55 Amylase Amylase activity activity (U/g koji) (U/g koji) 2 175 2 15 175 125 15 1 125 1 75 5 75 25 5 25,A a,,a A aa, b, A b,b ab,,b B aba, b, B b,bc ab,,bc B aab, b, B b,c b,,c B ba, b, B b 2.5 5 7.5 1 2.5 NaCl 5(%) 7.5 1,C b b,,c B ba, b c Temperature ( ) NaCl (%) Fig. 3. Effects of ph, temperature and NaCl concentration on the amylase activity of the rice koji with.5% laver. Symbols indicate:, ph 3;, ph 5;, ph 7;, ph 9. One unit of amylase activity was defined as the one milligram of maltose released from 1% soluble starch per a minute. The different capital letters indicate significantly different values in the horizontal column (P<.5). The different small letters indicate significantly different values in the vertical column (P<.5). Data expressed as the mean±sd. in triplicate. -amylase protease (Mikami et al., 1987; Siala 1 et al., 29). protease ph 1 9 8, 36 9 protease 7 8. 6 7, Protease Protease activity activity (U/g koji) (U/g koji) 5 6 4 5 3 4 2 3 1 2,E b, C b,d b,a c, A b,c c, B c ph, 1,A c,c b,c a. 1 9, A a, B a, C a, D a,,a A cb,,b B ba,,,b B bab, protease,c B b,,c B a 8 9,, A ca,, B ca,, C B ac,, C ca,, D ba, A b 48, B a, B ab, B b 7 8, A c, C c, C c, D b. Protease Protease activity activity (U/g koji) (U/g koji) 6 7 5 6 4 5 3 4 2 3 1 2
6 전준영ㆍ이미향ㆍ정인학ㆍ정민정ㆍ김병목 Protease activity (U/g koji) 1 9 8 7 6 5 4 3 2 1,E b, C b, D b,d b, C a,a c, A b, A b 15 25 35 45 55,C c, B c, AB b Protease activity (U/g koji) 1 9 8 7 6 5 4 3 2 1,A c, A b, A c b, C c,c b, B b, C c 2.5 5 7.5 1,C a, D a, D b Temperature ( ) NaCl (%) Fig. 4. Effects of ph, temperature and NaCl concentration on the protease activity of the rice koji with.5% laver. Symbols indicate:, ph 3;, ph 5;, ph 7;, ph 9. One unit of protease activity was defined as the one microgram of leucine released from.5% casein per a minute. The different capital letters indicate significantly different values in the horizontal column (P<.5). The different small letters indicate significantly different values in the vertical column (P<.5). Data expressed as the mean±sd. in triplicate. ph, 온도및 NaCl 농도가 amylase 활성에미치는영향 ph, NaCl,.5%, 48, ph, NaCl amylase (Fig. 3)., ph 3-9, 15-55 amylase ph, ph. ph 9 amylase ph. amylase, 15 ph 5 7 ph 3 9, 25, 45, 55 ph 3, 5, 7 (P<.5). NaCl -1% amylase ph NaCl 2.5%, NaCl 1%., amylase ph NaCl, ph 9 NaCl ph 5 7 (P<.5). ph, 온도및 NaCl 농도가 protease 활성에미치는영향 ph protease amylase (Fig. 4). Amylase, protease ph 9 ph 35-45,. protease, 15 ph 5 ph 3, 7, 9, 25 ph 3 ph 5, 7, 9 (P<.5). 35 45 ph 5 7 ph 3 9 (P<.5). NaCl protease ph 9 amylase. ph 9 NaCl., amylase protease ph 9,, 35.,. amylase protease, 1% NaCl. 사사 (E15644-2),. References AOAC. 25. AOAC official methods of analysis, 18th ed.
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