Kor J Fish Aquat Sci 43(6), 581-588 한수지, 43(6), 581-588, 2010 멍게껍질추출색소및 CLA (Conjugated Linoleic Acid) 가함유된사료를섭취한고등어 (Scomber japonicus) 의일반성분및지방산조성 박은정 김종태 1 강석중 2 최병대 경상대학교해양식품공학과 / 해양산업연구소, 1 경남특수사료, 2 경상대학교해양생명과학과 Fatty Acid Composition of Chub Mackerel (Scomber japonicus) Fed a Diet Fortified with CLA and Ascidian (Halocynthia roretzi) Tunic Extracts Eun Jung Park, Jong Tae Kim 1, Seok-Joong Kang 2 and Byeong-Dae Choi* Deptartment of Seafood Science and Technology / Institute of Marine Industry, Gyeongsnag National University, Tongyeong 650-160, Korea 1 Gyeongnam Special Feed Company, Goseong 638-962, Korea 2 Deptartment of Marine Life Science / Institute of Marine Industry, Gyeongsnag National University, Tongyeong 650-160, Korea To examine the functional properties of conjugated linoleic acid (CLA) and ascidian tunic extracts in fish, we compared mackerel fed ascidian tunic extract and CLA (CA25) and a control group. The daily growth index of CA25 was 1.92 compared to 1.86 in the control group. The viscerosomatic index of CA25 was 36.7% lower than that of the control group. After 8 weeks, the protein content decreased from 19.7 to 17.5% in the CA25 group. The ascidian tunic extract content in the viscera was much higher than in muscle (0.13 vs. 0.03 mg/100 g) after 8 weeks. At the start, the n-3 fatty acid content of the experimental fish was 25.2% in muscle and 23.7% in viscera. The CLA content in muscle in the CA25 group was 2.1% after 4 weeks and 2.3% after 8 weeks. By contrast, the CLA content in viscera did not change after 8 weeks. Key words: Mackerel, Conjugated linoleic acid (CLA), Ascidian tunic extracts, Growth rate, Fatty acid composition 서론고등어양식은 2003년이후고등어치어를채집하여연안가두리에서키우는양식방법이통영에서성공한이후고등어양식에종사하는어업인의수가꾸준히증가하고있는추세이다. 더욱이 2008년국가기관인경상남도수산자원연구소가우리나라최초로고등어대량인공종묘생산을성공한이래양식고등어산업화 5개년계획을적극추진중에있다. 해마다생산변동폭이많은고등어를안정적으로생산하여국민들에게공급하기위함이며무엇보다도양식고등어는장거리수송이가능한장점을갖고있다. 고등어양식이늘어나면서치사율이높아장거리유통이불가능하다는선례를깨고신선한횟감용으로고등어소비가가능해졌다. 고등어양식을통해풍부한필수지방산의신선한섭취가가능해짐에따라질높은건강을추구하는소비자의요구를반영하여고품질의기능성고등어의생산을모색할시기가되었다. 이미다기능성지질소재로알려진 CLA (conjugated linoleic acid) 가함유된사료를돼지, 소, 닭에게급이하여기능이향상된식품으로다양하게이용하고있다 (Kim, 1998). 어류양식산업의 CLA이용은아직까지다른식품에비해많지않지만 Corresponding author: bdchoi@gnu.ac.kr 감성돔, 우럭과같은양식어류를대상으로 CLA 와 carotenoids 를사료에첨가하여항산화효과, 체지방감소효과를검증한연구사례가보고되고있다 (Guo, 2010). CLA 는이미전세계적으로잘알려진 linoleic acid 의이중결합위치및기하학적이성체를총칭하는기능성물질로써많은연구결과에의하여항암효과, 체지방감소, 항산화효과, 항동맥경화, 항산화효과등에관한보고가많다 (Ip et al, 2002). 이후 CLA 는외국에서건강을위한기능성건강보조제로인증되었고, 우리나라역시 2006 년부터한국식품의약품안전청 (KFDA) 에서기능성식품원료사용을개별승인받았고, 2009 년에는고시형식품으로인정되기에이르렀다 (KFDA, 2009). Carotenoids 역시강력한항산화력을가진천연항산화제로써암발생억지, 어육의색상개선등의유용한기능성을나타내고있으며오늘날식음료첨가물, 의약용제, 사료첨가물등으로도다양하게이용되고있다 (Bauerfeind, 1981). 양식사료에 CLA 만첨가할경우어류의성장률이감소하거나체지방감소에대한우려가있다 (Twibell et al., 2001). 그러나 Choi et al (1996) 의무지개송어를대상으로한연구에의하면멍게껍질색소를사료에첨가하여급이하였을경우육의색소침착효과이외에성장이우수함을보여주고있으며, 또한 Torrissen (1986) 이대서양연어치어를대상으로한실험 581
582 박은정 김종태 강석중 최병대 112 에서는천연색소를첨가했을때인공색소첨가보다성장이빠르다고보고하고있다. 이에근거하여 CLA 와멍게껍질로부터추출한색소를동시에급이하여기능성어류를생산하는방법이가능하리라여겨진다. 따라서본연구에서는 CLA 와멍게껍질추출색소의기능을검증하기위해양식고등어사료에이들성분을첨가하여사육하였을때양식고등어의성장과기능성에어떠한영향을주는가에대하여검토하고자하였다. 재료및방법 실험어본실험에사용된고등어 (Scomber japonicus) 는우리나라남해안에위치한통영시산양면연안가두리양식장 ( 크기 5 5 7 m) 에서 2009 년 10 월 15 일부터 12 월 15 일까지총 8 주동안사육실험을실시하였다. 양식방법은한그룹은일반사료를먹인고등어 (Control) 와다른그룹은일반사료에멍게껍질추출색소및 CLA 가첨가된사료를먹인고등어 (CA25) 로나누어먹이를급이하였다. 급이방법은하루 2 회실시하였으며, 실험어의재료는 0 주, 4 주, 8 주총 3 회에걸쳐양식장에서직접채집하여실험실로이송한후, 양식고등어의두부를강타하여즉살하였고, 전체무게와내장의무게를각각측정한후, 육과내장으로구분하여각종실험을실시하였다. 실험사료 양식고등어의사료는경남고성군영현면에소재한 ( 주 ) 경남특수사료에서생산된어류용 EP (extruded pellet) 사료를급이하였고, 실험사료의조성은 Table 1 과같다. CLA 및멍게껍질추출색소첨가사료는다른영양성분의양은동일하게하였고, CA25 에는 Control 의어유량을줄인만큼 CLA 2.5% 그리고멍게껍질추출색소 1% 를첨가하여보충하였다. 실험사료에첨가한멍게껍질추출색소는가공부산물인멍게껍질일정량에 3 배량의아세톤을침지한후, 실온에방치하는방법으로두차례반복하여색소성분을충분히추출, 여과한후여과된아세톤추출물을회전진공증발농축기로 40 이하에서농축하여제조하였고, 전체사료중 1% 양을첨가하였다 (Choi et al., 1994). CLA 는 ( 주 )HK 바이오텍에서화학적으로합성된자체규격검사를통과한 CLA 를전체사료양의 2.5% 가되도록첨가하였다. 본실험에사용된 CLA 는 linoleic acid 로부터합성하여 c9,t11-cla 이성체와 t10,c12-cla 이성체가각각 48% 씩함유되어있었고그외이성체는미량으로함유되어있었다. CLA 농도를 2.5% 로결정한것은 Kang and Choi (1998) CLA 사료를급이하여잉어의성장을측정한연구결과를참고하였다. 어류의성장측정 어류의성장측정실험은실험전일하루절식후채취하였으며, 각실험구에서 6 마리씩채취하여어체의무게와내장의무게를측정하였다. 전체 8 주간급이한사료량과체중의증가량을기준으로사료효율, 일일성장율및내장지수를계산하였다 (Park et al., 2007). Table 1. Compositions of the experimental diets Ingredients Dietary treatments (g/kg) Control CA25 Fish meal 550 550 Soybean meal 70 70 Wheat flour 250 250 Yeast 10 10 Vitamin mix 1) 10 10 Mineral mix 2) 10 10 Soybean oil 40 40 Fish oil (squid liver) 60 19 CLA (purity 79%) 0 31 Carotenoids 3) 0 10 The values are mean±s.d. (n=3). 1 Vitamin mixture (mg kg -1 diet or IU): thiamin, 50 mg; riboflavin, 60 mg; calcium pantothenate, 200 mg; biotin 1 mg; folic acid 20 mg; pyridoxine, 40 mg; cyanocobalamimin, 0.05 mg; niacin, 250 mg; ascorbic acid, 1000 mg; inositol, 400 mg; retinyl acetate, 8000 IU; DL-cholecalciferol, 2400 IU; DL-alpha tocopherol acetate 300 IU; sodium menadione bisulphate, 5 mg. 2 Mineral mixture (mg kg -1 ): calcium carbonate, 850 mg; magnessium oxide, 750 mg; copper sulphate, 25 mg; manganese sulphate, 100 mg; ferric citrate, 150 mg; zinc sulphate, 120 mg. 3 Carotenoids; Ascidian tunic extracts. 일반성분분석일반성분의분석은일정기간사육된양식고등어를육과내장으로나눈후, 일반성분은 AOAC 법 (1990) 에준하여수분은 105 에서상압가열건조법, 조회분은 550 에서건식회화법, 조단백질은 semi-micro Kjeldahl 법으로질소측정후단백질계수 (6.25) 를이용하였으며, 조지방은 Bligh and Dyer 법 (1959) 으로분석하였다. 총지질의추출과지방산조성지질의추출은 Bligh and Dyer 법 (1959) 에따라 chloroform: methanol 혼합용매 (2:1, v/v) 로반복추출한다음포화식염수을이용하여지질과수분을분리한후, 지질층을농축하여클로로포름 100 ml 로정용한다음무게를측정하여정량하였다. 지방산조성분석을위해 AOCS 법 (1990) 에따라 50 mg 의지질시료를 cap tube 에넣고 0.5 N NaOH-methanol 용액을더하여질소충전한후, 100 에서 3 분간가열하여검화하였다. 방냉후 1.0 N H 2SO 4 2 ml 를더한후, 55 에서 20 분간가열하여 methylester 화하였다. 이를상온에냉각한후 isooctane 1 ml 를첨가한후 30 초간 vortex mixer 로혼합하였다. 이후 5 ml 의포화식염수를더하여흔들어방치한후 isooctane 층이분리되도록하였다. 분리된 isooctane 층은시료병에옮고 cap tube 에다시 isooctane 1 ml 를더하여재추출하여시료병에모았다. 이를 capillary column (Omegawax-320, 30 m 0.25 mm i.d., Supelco Co., Bellefonte, PA, USA) 이장착된 gas chromatography (Shimadzu GC-17A, Shimadzu Co, Kyoto, Japan) 을이용하여분석하였다. 이때의분석조건은 column 온
127 멍게껍질추출색소및 CLA 가함유된사료를섭취한고등어 (Somber Japonicus) 의일반성분및지방산조성 583 도 180 에서 5 분간유지후 230 까지 3 /min 씩온도를올려 15 분간유지하였고, injector 온도 250, detector 온도 26 0, 그리고 carrier gas 는 He (1.0 kg/cm 2 ) 을사용하였으며 split rate 는 1:100 으로설정하였다. 각구성지방산의동정은표품과의머무름시간 (RT) 의비교및 equivalent chain length 법에의해동정하였다. Carotenoids 함량의측정총 carotenoids 의함량은 460 nm 에서 Umeda et al. (1971) 의방법에따라 UV/VIS spectrophotometer (Shimadzu UV-1700, Shimadzu Co., Kyoto, Japan) 로측정하고다음의식에의하여계산하였다. OD (λmax) Vol 1,000 Total carotenoids (mg/100 g) = E 1% 1cm (2,660) weight sample (g) 이때 E 1% 1cm 는 extinction coefficient 를나타내며, 시료의용매 chloroform 의흡광계수 2,660 을사용하였다 (Whang, 1999). 통계처리각실험은모두 3 회이상반복실험을통해결과를얻었으며, 결과의통계처리는 SPSS (Statistical package for the social science, Chicago Illinois USA) program version 12.0 을이용하였으며, 각각의시료에대해서는평균 ± 표준편차로나타냈다. 각시료에대한유의차검정은 One-way ANOVA test 와 T-test 를실시하여분산분석한후, Duncan's multiple range test 에의해정직유의차검정을실시하여유의차검정 (P<0.05) 을실시하였다. 결과및고찰 사료의일반성분및지방산조성의변화시판어분을기본으로하여미네랄및비타민을첨가하였고, 첨가된어유는 6% 를기본으로하여 CA25 구에는 CLA 가 2.5% 함유되도록하여전체적으로는첨가되는지질의함량이일정하도록하였다 (Table 1). 제조된사료의수분함량은 27.0-27.6%, 단백질함량은 31.9-32.3%, 지질함량은 5.8-7.3%, 회분은 8.1-8.8% 였다. 사료의성분조성은 Table 2 와같다. Table 2. Proximate composition of the experimental diets Proximate composition Dietary treatments (%) Control CA25 Moisture 27.6±0.3 a 27.0±0.1 b Crude protein 32.3±5.1 a 31.9±1.6 b Crude lipid 5.8±0.5 b 7.3±0.5 a Ash 8.8±0.2 a 8.1±0.2 b Table 3. Fatty acid compositions of experimental diets (% of total fatty acids) 14:00 3.9 ± 0.0 3.4 ±0.2 15:00 0.5 ± 0.0 0.4 ±0.0 16:00 16.1 ± 0.1 14.2 ±0.0 18:00 4.5 ± 0.1 4.0 ±0.0 Saturates 26.8 24.0 16:1n-7 5.0 ± 0.1 4.4 ±0.0 18:1n-9 18.9 ± 0.2 17.2 ±0.1 18:1n-7 3.0 ± 0.1 2.6 ±0.0 20:1n-11 0.9 ± 0.0 1.4 ±0.1 20:1n-9 1.3 ± 0.9 1.6 ±0.0 22:1n-11+13 1.7 ± 0.0 1.3 ±0.0 22:1n-9 0.4 ± 0.0 0.4 ±0.0 Monoenes 32.1 29.0 16:2n-4 0.5 ± 0.0 0.5 ±0.0 16:3n-4 0.7 ± 0.1 0.8 ±0.1 16:4n-1 0.9 ± 0.0 1.2 ±0.0 18:2n-6 15.2 ± 0.1 12.1 ±0.2 18:3n-3 2.0 ± 0.2 1.4 ±0.0 CLA-1 1) 0.0± 0.0 6.5 ±0.1 18:4n-3 1.3 ± 0.0 1.2 ±0.0 CLA-2 2) 0.0 ± 0.0 6.7 ±0.1 20:4n-6 0.8 ± 0.1 0.6 ±0.0 20:5n-3 6.0 ± 0.1 5.8 ±0.1 22:5n-3 1.6 ± 0.1 1.4 ±0.0 22:6n-3 7.7 ± 0.6 5.8 ±0.0 Polynenes 42.0 48.0 Total CLA 0.0 13.1 n-6 17.0 a 13.3 b n-3 19.9 a 16.5 a n-3/ n-6 1.17 a 1.24 a 1 CLA-1: c9,t11-cla, 2 CLA-2: t10,c12-cla. 어류사료에포함된지방산의종류와함량은어류의성장에큰영향을미칠수있으므로각어종의필수지방산을고려하여어유의첨가량과종류를결정하는것이바람직하다. Covadonga et al. (2004) 이실험한 양식돔과자연산돔의지방산조성실험 을위해제조된사료조성을근거해볼때고등어의정상적인성장을위해본실험의사료는지방산조성이알맞게분포되어있었다 (Table 3). 포화지방산의함량은 24.0-26.8%, 모노엔산의함량은 29.0-32.1%, 그리고다가불포화산의함량은 42.0-48.0% 로풍부하였다. 다가불포화산중 C18:2n-6 의함량이높은것은사료에첨가된대두유의영향으로여겨지며, CA25 군의경우 C20:5n-3 과 C22:6n-3 의함량이약간낮은것은 CLA 의증가로인해상대적으로낮게나타난것으로생각된다. 그러나 Σn-3/Σn-6 비를계산한결과 Control 구의비는 1.17, CA25 구의비는 1.24 로 CLA 의영향으로 n-3 의함량이낮아 Σn-3/Σn-6 비는약간높게나타났다. CA25 구의 cis-9, trans-11 CLA 는 6.5%, trans-10, cis-12 CLA 는 6.7% 로고른분포를나타내고있었다.
584 박은정 김종태 강석중 최병대 112 Table 4. Growth performance of mackerel fed with different diets for 8 weeks Control CA25 Initial body weight (g) 272.0±22.6 a 272.0±22.6 a Final body weight (g) 423.3±25.8 b 431.3±27.4 a Initial viscera weight (g) 26.0±5.1 a 26.0±5.1 a Final viscera weight (g) 44.8±6.8 a 37.5±3.9 b DGI 1) 1.86 a 1.92 a FCR 2) 1.00 a 1.00 a VSI 3) 12.0 a 7.60 b The values are mean±s.d. (n=6). Different superscript letters 1 DGI; Daily growth index=100 [(Final body weight) 1/3 - (Initial body weight) 1/3 ]/days. 2 FCR; Feed conversion ratio=100 [Total fed amount/(final body weight-initial body weight)]. 3 VSI; Viscerosomatic index=100 (viscera weight/body weight). 실험기간중의고등어성장실험사료를급이한고등어성장에관한자료는 Table 4 와같다. 급이를시작할시점의무게는 272.0 g 이었다. 급이 8 주후 Control 구는 423.3±25.8 g, CA25 구는 431.3±27.4 g 이었다. 늦가을의낮은수온이였음에도불구하고체중만으로평가하면오히려 CA25 구의체중이더많이늘고, 내장무게의절대증가분을보면 CA25 구경우전체고등어의무게중내장의함량이 Control 구에비해 28% 나적어상대적으로육의함량이증가되었음을알수있었다. 내장계수 (VSI) 를보면 Control 구의 63% 에불과함을알수있어단순히체중증가뿐아니라육의함량이매우증가되었음을알수있었다. 더욱이두실험구간에사료효율은같은조건에서 Control 구와 CA25 구가같은 1.00 을나타내었다. 어종에따라다르겠지만본실험에서는일반사료와 CLA 를첨가한사료와비교했을때사료효율이나일일성장율에는서로간에큰영향을미치지않았다. Bandarra et al. (2006) 은 CLA 를급이하여사육한송어내장지방의무게가감소하는것을보고하고있으며, Kennedya et al. (2007) 은대구에 fish oil 과 CLA 를급이하여각각사육하였는데, CLA 를급이한대구의내장계수가더작으며사료효율역시차이가없음을보고하고있다. 고등어의일반성분변화 8 주간사료를급이한후측정한고등어의일반성분함량은 Table 5 에나타내었다. 고등어육의수분함량은처음엔 65.1% 였지만, 4 주이후 8% 가까이감소하여 57.0-58.0% 를보였으며, 그이후 8 주때는 54.8-57.0% 로나타났다. 이는같은기간동안지질의증가에기인된것으로여겨진다 (23.1-24.3%). 단백질함량의경우처음 18.0% 가 4 주에는 19.7-21.2% 로소량증가하였지만, 8 주후에는 17.4-17.5% 로오히려실험시작시점보다감소하는것으로나타났다. 그러나 4 주 Control 18.6-19.1%, 4 주 CA25 19.2-21.1%, 8 주 Control 23.1-32.1%, CA25 24.3-26.5% 로현저히증가함을보여준다. 이는감성돔을대상 으로같은내용의실험을한 Guo (2009) 의연구결과와도같은경향을보여주고있다. 다만 Jeong et al. (1998) 의연구결과에의하면계절적으로 11 월이후지질의함량이감소해야함에도불구하고실험기간중두배가까이증가를보이는것은, 산란회유시에너지원및생식선의성숙을위해지질이소모되어야하는데회유하지못하고가두리에갇혀활동량이현저히줄어들어지방의축적된것으로추정되었다. CLA 의양은 CLA 가첨가된사료를먹인 4 주및 8 주후 CA25 구각각의육에서는전체지방산중에서 2.1%, 2.3% 로나타났지만내장에서는모두전체지방산중 1.8% 로나타났다. 이는 Guo (2009) 가실험한감성돔과우럭의실험결과에비추어볼때, 고등어에 CLA 가축적된양이같은기간에비해상대적으로적음을알수있었다. 회분의함량은 1.3-1.8% 로모두적은양이었다. Table 5. Proximate composition (%) of the mackerel fed with different diets for 8 weeks 0 week 4 week 8 week Control Control CA25 Control CA25 Moisture 65.1±3.1 58.0±0.5 57.0±0.4 57.0±0.8 54.8±0.8 Protein 18.0±2.3 21.2±3.0 19.7±0.9 17.4±1.0 17.5±2.5 (51.6) (50.4) (45.8) (40.4) (38.7) 1) 11.1±2.6 18.6±0.3 19.2±0.9 23.1±2.4 24.3±0.1 Lipid M (31.8) (44.3) (44.7) (51.7) (54.6) 2) 14.1±2.6 19.1±3.2 21.1±0.2 32.1±2.7 26.5±3.1 Lipid V (40.4) (51.7) (49.1) (71.8) (59.6) CLA M 2.1 2.3 CLA V 1.8 1.8 Ash 1.6±0.4 (3.6) 1.3±0.4 (2.9) 1.8±0.2 (4.0) 1.4±0.0 (3.1) 1.3±0.3 (2.9) The values are mean±s.d. (n=3). Parenthesis is dry basis. 1 M; Muscle, 2 V; Viscera. Table 6. Carotenoids content of muscle and viscera in mackerel fed with different diets for 8 weeks Feeding Dietary treatments (mg/100 g) Samples periods Control CA25 4 week Muscle 0.01±0.0 b 0.08±0.0 a Viscera 0.02±0.1 b 0.15±0.0 a 8 week Muscle 0.03±0.0 b 0.10±0.0 a Viscera 0.03±0.0 b 0.13±0.0 a 고등어육과내장에서의 carotenoids 함량변화고등어사육 4 주, 8 주후육과내장의 carotenoids 함량은 Table 6 에나타내었다. 본실험에서는멍게껍질에서추출한 carotenoids 를사용하였는데이는 Lee et al. (1994) 에의해멍게껍질추출물이합성첨가제인 carophyll pink (Hoffman La-Roche, Basle, Switzerland) 와같은착색효과가있음이입증되었기때문이다. Carotenoids 는발색의원인이되는공액이중결합이중복된장쇄상의구조를가지고있어생리적으로생체
127 멍게껍질추출색소및 CLA 가함유된사료를섭취한고등어 (Somber Japonicus) 의일반성분및지방산조성 585 에유해한자유라디칼 (free radical) 을포집하거나뛰어난항산화기능을나타낸다 (Tee, 1992). 특히 β-carotene 과 astaxanthin 등은강력한항산화효과뿐아니라성장률개선, 질병발생억제, 어육의색상개선등의기능성을나타낸다고보고되고있으며 (Hong et al., 1998), 일반적으로 carotenoids 는지용성이며공액이중결합의수가많을수록황색에서적색으로이행하는것으로알려져왔다 (DuPlessis and Grobbelaar, 1979). 실험결과육의경우 CA25 구에서 4 주, 8 주모두 0.08±0.0 mg/100 g, 0.10±0.0 mg/100 g 그리고내장의경우는 0.15±0.0 mg/100 g, 0.13±0.0 mg/100 g 으로육에비해내장의축적율이약간높긴하지만모두그리높은축적율을나타내지는않았다. 이는 Kang and Choi (2009) 가무지개송어의발색을위해 carophyll pink 와멍게껍질추출물 (ascidian tunic extracts) 을각각급이하였을때 carophyll pink 첨가구에비해멍게껍질추출물첨가구의함량이낮다고보고하고있는데어류에따른 carotenoids 대사의차이로보인다. Table 7. Fatty acid compositions of muscle and viscera lipid in mackerel at the start of the experiment (% of total fatty acid) Fatty acids Muscle Viscera 14:0 3.6 ± 0.0 3.7 ± 0.5 15:0 0.4 ± 0.0 0.4 ± 0.1 16:0 17.1 ± 0.0 19.3 ± 0.8 17:0 0.5 ± 0.4 0.5 ± 0.1 18:0 4.8 ± 1.1 +5.2 ± 0.1 Saturates 27.1 29.7 16:1n-7 6.3 ± 0.1 6.4 ± 0.6 18:1n-9 19.3 ± 0.1 19.8 ± 0.6 18:1n-7 3.9 ± 0.1 3.9 ± 0.2 20:1n-11+9 2.5 ± 0.0 2.1 ± 0.8 22:1n-11+13 1.6 ± 0.1 1.5 ± 0.5 22:1n-9 0.5 ± 0.1 0.6 ± 0.3 Monoenes 35.6 36.1 16:2n-4 1.1 ± 0.0 1.0 ± 0.1 16:3n-4 0.7 ± 0.0 0.9 ± 0.1 16:4n-1 0.6 ± 0.1 0.4 ± 0.1 18:2n-6 1.7 ± 0.1 1.3 ± 0.5 18:3n-4 0.5 ± 0.4 0.2 ± 0.0 18:3n-3 0.8 ± 0.1 0.6 ± 0.1 18:4n-3 1.7 ± 0.0 1.4 ± 0.0 20:2NMID 0.4 ± 0.1 0.4 ± 0.1 20:4n-6 1.2 ± 0.0 1.2 ± 0.0 20:4n-3 0.6 ± 0.5 0.5 ± 0.1 20:5n-3 7.6 ± 0.0 7.7 ± 0.6 21:5n-3 0.5 ± 0.1 0.5 ± 0.2 22:5n-3 1.8 ± 0.6 1.9 ± 0.3 22:6n-3 11.7 ± 0.0 10.9 ± 0.5 Polyenes 34.5 31.6 Total CLA 0.0 0.0 n-6 4.4 a 3.5 a n-3 25.2 a 23.7 a n-3/ n-6 5.7 a 6.7 a 실험초기고등어의육과내장의지방산조성실험의시작전사육된고등어육과내장의지방산조성을 Table 7 에나타내었다. 어류지질을구성하고있는지방산조성의변화를보면육의경우 C16:0 과 C18:1 은각각 17.1% 및 23.2% 였고, EPA (C20:5n-3) 조성비는 7.6%, DHA (C22:6n-3) 조성비는 11.7% 로나타났다. 내장의경우 C16:0 과 C18:1 은각각 19.3% 및 23.7% 였고, EPA 조성비는 7.7%, DHA 조성비는 10.9% 로육과내장의지방산조성에큰차이는없었다. 또한육과내장모노엔산은각각 35.6%, 36.1%, 폴리엔산은 34.5%, 31.6%, n-3 HUFA 조성비는 25.2%, 23.7% 로함량에있어그리큰차이를보이지않았다. 특히고등어의 HUFA 의경우전체지방산중 31.6-34.5% 를차지하는데이중 n-3 HUFA 가차지하는비중이 73.0-75.0% 이었다. Table 8. Fatty acid compositions of muscle lipid in mackerel after feeding the experimental diets for 4 week (% of total fatty acid) 14:0 2.9 ± 0.5 3.1 ± 0.2 16:0 19.0 ± 0.6 18.6 ± 0.4 17:0 0.5 ± 0.1 0.5 ± 0.1 18:0 5.6 ± 0.2 6.9 ± 0.4 Saturates 28.9 30.0 16:1n-7 5.8 ± 0.2 5.6 ± 0.3 18:1n-9 25.7 ± 0.8 23.2 ± 0.3 18:1n-7 4.1 ± 0.1 3.8 ± 0.2 20:1n-11+9 2.4 ± 0.6 3.1 ± 0.5 22:1n-11+13 1.4 ± 0.4 1.4 ± 0.3 Monoenes 40.8 38.5 16:2n-4 1.0 ± 0.1 0.6 ± 0.5 16:3n-4 0.9 ± 0.1 0.8 ± 0.0 18:2n-6 3.0 ± 0.5 3.3 ± 0.8 18:3n-6 0.3 ± 0.0 0.3 ± 0.0 18:3n-4 0.3 ± 0.0 0.2 ± 0.1 18:3n-3 0.7 ± 0.0 0.6 ± 0.1 CLA-1 1) 0.0 ± 0.0 1.1 ±+0.5 18:4n-3 1.1 ± 0.1 1.1 ± 0.1 CLA-2 2) 0.0 ± 0.0 1.0 ± 0.5 20:2NMID 0.6 ± 0.1 0.5 ± 0.1 20:4n-6 1.3 ± 0.1 0.8 ± 0.5 20:4n-3 0.5 ± 0.0 0.5 ± 0.0 20:5n-3 5.6 ± 0.2 5.7 ± 0.2 21:5n-3 0.3 ± 0.0 0.3 ± 0.0 22:4n-6 0.4 ± 0.0 0.3 ± 0.1 22:5n-3 1.8 ± 0.1 1.8 ± 0.1 22:6n-3 9.7 ± 1.0 9.2 ± 0.9 Polyenes 29.6 30.5 Total CLA 0.0 2.1 n-6 5.6 a 5.3 a n-3 20.0 a 19.4 a n-3/ n-6 3.6 a 3.6 a 1 CLA-1; c9,t11-cla, 2 CLA-2; t10,c12-cla.
586 박은정 김종태 강석중 최병대 112 Table 9. Fatty acid compositions of viscera lipid in mackerel after feeding the experimental diets for 4 week (% of total fatty acid) 14:0 3.0 ± 0.3 3.0 ± 0.1 15:0 0.4 ± 0.1 0.4 ± 0.1 16:0 17.8 ± 1.7 18.2 ± 0.7 17:0 0.5 ± 0.1 0.4 ± 0.1 18:0 8.1 ± 3.8 7.1 ± 0.4 Saturates 30.3 29.7 16:1n-7 5.7 ± 0.4 5.5 ± 0.2 18:1n-9 25.3 ± 1.2 24.2 ± 1.6 18:1n-7 4.2 ± 0.4 3.9 ± 0.2 18:1n-5 0.3 ± 0.1 0.3 ± 0.1 20:1n-11+9 3.3 ± 0.7 3.0 ± 0.5 22:1n-11+13 1.7 ± 0.4 1.7 ± 0.6 Monoenes 41.6 39.9 16:2n-4 1.1 ± 0.1 1.0 ± 0.1 16:3n-4 0.9 ± 0.2 0.8 ± 0.1 18:2n-6 2.7 ± 0.6 3.0 ± 0.4 18:3n-6 0.4 ± 0.3 0.2 ± 0.0 18:3n-3 0.6 ± 0.0 2.2 ± 2.8 CLA-1 1) 0.0 ± 0.0 0.9 ± 0.2 18:4n-3 1.1 ± 0.2 0.9 ± 0.0 CLA-2 2) 0.0 ± 0.0 0.8 ± 0.2 20:4n-6 1.5 ± 0.0 1.3 ± 0.1 20:4n-3 0.6 ± 0.1 0.6 ± 0.3 20:5n-3 5.6 ± 0.7 5.3 ± 0.4 22:4n-6 0.3 ± 0.0 0.3 ± 0.0 22:5n-3 1.7 ± 0.2 1.5 ± 0.1 22:6n-3 7.4 ± 3.6 8.3 ± 0.9 Polyenes 27.5 30.2 Total CLA 0 1.8 n-6 5.7 a 5.4 a n-3 17.5 a 19.2 a n-3/ n-6 3.1 a 3.6 a The values are mean±s.d.(n=3). Different superscript letters 1 CLA-1; c9,t11-cla, 2 CLA-2; t10,c12-cla. 고등어사육기간중육및내장지방산조성의변화 CLA 가함유된사료와함유되지않은사료를각각 8 주동안고등어에급이하면서체내축적량및내장중축적량의변화를 Table 8, 9, 10 및 11 에각각나타내었다. 육의포화지방산의조성비는 4 주후 Control 구, CA25 구에서각각 28.9%, 30.0% 였으며, 모노엔산의조성비는각각 40.8%, 38.5% 였고, 폴리엔산의조성비는각각 29.6%, 30.5% 로나타났다. 지방산의전체조성은큰변화를나타내지않았으나, 다만 CA25 군에서 CLA 가 2.1% 축적되었고 CLA-1 과 2 사이에양의차이는거의없었다. 양식 8 주후육의포화지방산의조성비는 Control 구, CA25 구에서각각 27.5%, 28.8%, 모노엔산의조성비는각각 44.6%, 41.0%, 폴리엔산의조성비는각각 29.4%, 31.9% 로나타났고, 4 주와 8 주의급이기간으로인한지방산조성의큰변화는없었다. 역시 CA25 의경우에는 CLA 가 2.3% 축적되었고, CLA-1 과 2 사이에함량의차이도거의없었다. 실험 8 주간의사료급이기간동안 CA25 구에서 n-6 계지방산의증가를보이고있는데 이는각각의사료에대두유를첨가한것이그원인으로추정되었다. 그러나초기에는 5.7 이었던 n-3/ n-6 의비가 4 주에 3.6, 8 주에 3.3 정도에머무는것으로보아 n-3 필수지방산의추가급이는하지않더라도성장에는영향을미치지않는것으로여겨진다. Table 10. Fatty acid compositions of muscle lipid in mackerel after feeding the experimental diets for 8 week (% of total fatty acid) 14:0 2.9 ± 0.1 2.9 ± 0.0 15:0 0.3 ± 0.0 0.3 ± 0.0 16:0 18.1 ± 0.5 17.9 ± 0.0 17:0 0.4 ± 0.0 0.4 ± 0.0 18:0 5.4 ± 0.1 6.7 ± 0.1 20:0 0.2 ± 0.0 0.3 ± 0.0 Saturates 27.5 28.8 16:1n-7 6.0 ± 0.1 5.7 ± 0.1 18:1n-9 28.2 ± 0.5 25.4 ± 0.1 18:1n-7 4.4 ± 0.1 4.0 ± 0.0 20:1n-11 1.1 ± 0.3 0.9 ± 0.0 20:1n-9 2.2 ± 0.4 2.1 ± 0.1 22:1n-11+13 1.6 ± 0.1 1.8 ± 0.1 Monoenes 44.6 41.0 16:2n-4 0.3 ± 0.0 0.4 ± 0.0 16:3n-4 0.8 ± 0.0 0.7 ± 0.0 18:2n-6 3.1 ± 0.2 3.5 ± 0.1 18:3n-3 0.7 ± 0.0 0.7 ± 0.0 CLA-1 1) 0.0 ± 0.0 1.2 ± 0.2 18:4n-3 0.9 ± 0.0 1.0 ± 0.0 CLA-2 2) 0.0 ± 0.0 1.1 ± 0.2 20:2NMID 0.6 ± 0.1 0.5 ± 0.0 20:4n-6 1.3 ± 0.1 1.2 ± 0.0 20:3n-3 0.3 ± 0.0 0.0 ± 0.0 20:4n-3 0.5 ± 0.0 0.5 ± 0.0 20:5n-3 5.5 ± 0.1 5.8 ± 0.1 22:5n-3 1.7 ± 0.1 1.7 ± 0.0 22:6n-3 9.0 ± 0.3 8.9 ± 0.3 Polyenes 29.4 31.9 Total CLA 0.0 2.3 n-6 5.6 a 5.7 a n-3 19.0 a 19.0 a n-3/ n-6 3.4 a 3.3 a The values are mean±s.d.(n=3). Different superscript letters 1 CLA-1; c9,t11-cla, 2 CLA-2; t10,c12-cla. Table 11. Fatty acid compositions of viscera lipid in mackerel after feeding the experimental diets for 8 week (% of total fatty acid) 14:0 2.6 ± 0.1 2.6 ± 0.2 16:0 17.5 ± 0.4 17.4 ± 0.5 17:0 0.4 ± 0.0 0.4 ± 0.0 18:0 5.7 ± 0.2 7.1 ± 0.4 Saturates 27.0 28.5 16:1n-7 5.2 ± 0.3 5.6 ± 1.3 18:1n-9 29.0 ± 0.3 26.8 ± 1.4 18:1n-7 4.5 ± 0.0 4.2 ± 0.2 20:1n-11 0.7 ± 0.0 0.8 ± 0.2 20:1n-9 2.5 ± 0.1 2.2 ± 0.1 22:1n-11+13 1.5 ± 0.1 1.5 ± 0.2
127 멍게껍질추출색소및 CLA 가함유된사료를섭취한고등어 (Somber Japonicus) 의일반성분및지방산조성 587 Table 11. continued Monoenes 44.7 42.0 16:3n-4 0.7 ± 0.0 0.7 ± 0.0 18:2n-6 2.6 ± 0.2 2.8 ± 0.3 18:3n-3 0.6 ± 0.0 0.6 ± 0.0 CLA-1 1) 0.0 ± 0.0 1.0 ± 0.2 18:4n-3 1.1 ± 0.3 0.9 ± 0.1 CLA-2 2) 0.0 ± 0.0 0.8 ± 0.2 20:2NMID 0.8 ± 0.3 0.7 ± 0.2 20:4n-6 1.4 ± 0.1 1.3 ± 0.1 20:4n-3 0.6 ± 0.1 0.5 ± 0.0 20:5n-3 5.4 ± 0.2 5.6 ± 0.3 22:5n-3 1.7 ± 0.1 1.7 ± 0.0 22:6n-3 9.3 ± 0.1 9.1 ± 0.4 Polyenes 29.8 30.7 Total CLA 0.0 1.8 n-6 5.3 a 5.2 a n-3 19.3 a 18.8 a n-3/ n-6 3.6 a 3.6 a The values are mean±s.d.(n=3). Different superscript letters 1 CLA-1; c9,t11-cla, 2 CLA-2; t10,c12-cla. 양식 4 주후내장의포화지방산의조성비는 Control 구, CA25 구에서각각 30.3%, 29.7%, 모노엔산의조성비는각각 41.6%, 39.9%, 폴리엔산의조성비는각각 27.5%, 30.2% 로각군간의큰차이를보이지않았다. 양식 4 주동안내장에 CLA 는 1.8% 축적되었으며, CLA-1 과 2 사이에양의차이는거의없었다. 양식 8 주후내장의포화지방산의조성비는 Control 구, CA25 구에서각각 27.0%, 28.5%, 모노엔산의조성비는각각 44.7%, 42.0%, 폴리엔산의조성비는각각 29.8%, 30.7% 로나타나육과마찬가지로 4 주와 8 주사료급이를통한급격한지방산의변화는없었다. CLA 역시 4 주와같은 1.8% 가축적이되었고, CLA-1 과 2 사이에양의차이는거의없었다. 실험초기 6.7 이었던 n-3/ n-6 의비 4 주, 8 주모두 3.6 에머무는것으로보아육과마찬가지로식물성사료로인해 n-6 지방산이초기에증가하기는하였지만추가급이를하더라도변동이없음을알수있었다. 4 주와 8 주, 육과내장의두차례에걸친실험을통해사료급이후 CLA 의축적량과 n-6 계지방산증가량을보면 8 주이상의추가급이를통해변화를기대하기는어려울것으로추측되었다. Schut et al. (1997) 에의하면쥐등의설치류를대상으로한 CLA 생리활성실험에서체내에흡수된 CLA 가모노엔산조성비를약간감소시킨다고보고하고있는데, 이는본실험의결과역시성장하면서전체적인 monoenes 의함량이증가하지만, 4 주급이후 Control 구와 CA25 군의육에서는각각 40.8%, 38.5%, 내장에서는각각 41.6%, 39.9% 로 CA25 구가 Control 구에비해 monoenes 의조성비가낮음을알수있었다. 이는 8 주후실험에도동일한결과를보여 Control, CA25 육에서도각각 44.6%, 41.0%, 내장에서도각각 44.7%, 42.0% 를나타내고있었다. CLA 가다양한대사를통해동물체내에서항암, 항동맥경화, 항비만등의탁월한효과가입증되어기능 성식품으로인정을받았고, 실제로사료에 CLA 를첨가하여기능성식품을생산하는추세가늘었음에도불구하고 (Ha et al., 1998) 양식어민들이사료에 CLA 첨가를우려하는것은 CLA 가어류의지방축적을억제시킨다는연구결과때문이다 (Azain et al., 2000). 그러나 Twibell et al. (2001) 은어류실험을통해 CLA 수준이증가할때성장률이약간감소하였지만사료효율은오히려향상되었다고보고하고있다. 그러므로본실험의결과로볼때 CLA 와같은기능성사료급이의시기와양을적절히조절할경우기능성어류의생산을극대화하는효율적인기준을만들수있을것으로여겨진다. 사 본연구과제는 2009 년도경남테크노파크에서시행한지역기반육성기술개발사업연구비지원에의해수행된결과의일부이며, 이에감사드립니다 ( 과제번호 : 20094001). 참고문헌 Ackman RG. 1995. Composition and nutritive value of fish and shellfish lipids. In: Fish and Fishery Products. Ruither A, ed. CAB International, London, U.K., 117-156. AOAC. 1995. Official Methods of Analysis, 16th ed. Association of Official Analytical Chemists, Washington DC, U.S.A., 69-74. AOCS. 1990. AOCS official Methods of Ce 1b-89. In: Official Methods and Recommended Practice of the AOCS. 4th ed., Association of Official Chemists and Science, Champaign, IL, U.S.A. Azain MJ, Hasuman DB, Sisk MB, Flatt WP and Jewell DE. 2000. Dietary conjugated linoleic acid reduces rat adipose tissue cell size rather than cell number. J Nutrition 130, 1548-1554. Bandarra NM, Nunes ML, Andrade AM, Prates JAM, Pereira S, Monateiro M, Rema P and Valente LMP. 2006. Effect of dietary conjugated linoleic acid on muscle, liver and visceral lipid deposition in rainbow trout juveniles (Oncorhynchus mykiss). Aquaculture 254, 496-505. Bauerfeind JC. 1981. Natural food color. In: Carotenoids as Colorants and Vitamin A Precursors. Bauerfeind JC, ed. Academy Press Inc, New York, U.S.A. 1-47. Bligh EG and Dyer WT. 1959. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917. Choi BD, Kang SJ and Lee KH. 1996. Quality improvement of rainbow trout with pigments and enzymatic hydrolysates of ascidian (Halocynthia roretzi) tunic 2. Effects of ascidian tunic enzymatic 사
588 박은정 김종태 강석중 최병대 112 hydrolysates on pigmentation and growth of rainbow trout (Oncorhynchus mykiss). J Korean Fish Sci 29, 359-368. Choi BD, Kang SJ, Choi YJ, Youm MG and Lee KH. 1994. Utilization of ascidian (Halocynthia roretzi) tunic 3. Carotenoid compositions of ascidian tunic. Bull Korean Fish Soc 27, 344-350. Covadonga R, Carolina A, Pilar B, Juana RC, Francisco JS and Antonio I. 2004. Assential of lipid essential fatty acids requirements of black seabream (Spondyliosome cantharus) by comparison of lipid composition in muscle and liver of wild and capitive adult fish. Com Biochem Physio Part B 139, 619-629. DuPlessis LM and Grobbelaar N. 1979. Isomerization of the double bonds of a conjugated fatty acid during beta-oxidation. Lipids 14, 943-948. Guo R. 2010. Conjugated linoleic acid (CLA) deposition and antioxidant activity of lipid extracted from black seabream (Acanthopagrus schlegeli) and rockfish (Sebastes schlegeli) fed with CLA and carotenoids. M.S. Thesis, Gyeongsang National University, Tongyeong, Korea. Ha YL, Park GB, Kang SJ, Park WS and Park KA. 1998. Development and utilization of prospective biofunctional resources in agriculture. In: Proceeding 1st Agr. Res. Util. Symposium for 50th Anniversary. Ha YL, ed. Gyeongsang National University, Jinju, Korea, 49-55. Hong SP, Kim MH and Whang JK. 1998. Biological functions and production technology of carotenoids. Korean Soc Food Sci Nutr 27, 1297-1306. Ip C, Dong Y, Ip MM, Banni S, Carta G, Angioni E, Murru E, Spada S, Melis MP and Saebo A. 2002. Conjugated linoleic acid isomers and mammary cancer prevention. Nutr Cancer 43, 52-58. Jeong BY, Moon SK, Choi BD and Lee JS 1998. Seasonal variation in lipid class and fatty acid composition of 12 species of Korean fish. J Korea Fish Soc 32, 30-36. Kang SJ and Choi BD. 1997. Encouragement of cooperative research after openness of import fishes/production of fish containing newly recognized anticarcinogen CLA(Conjugated Linoleic Acid). J Ins Marin Industry 8, 3-17. Kang SJ and Choi BD. 2009. Composion in carotenoids of rainbow trout fed with CLA and ascidian tunic extracts. Kor J Fish Aqust Sci 42, 721-724. Kennedya SR, Bickerdikeb R, Bergec RK, Portera AR and Tochera DR. 2007. Influence of dietary conjugated linoleic acid (CLA) and tetradecylthioacetic acid (TTA) on growth, lipid composition and key enzymes of fatty acid oxidation in liver and muscle of Atlantic cod (Gadus morhua L.). Aquaculture 264, 372-382. KFDA, 2009. Korea Food & Drug Adminastration. http://hfoodi.kfda.go.kr/ material/ Kim JO. 1998. Newly recognized multifunctional fatty acids for the production of high quality meat, fish and agricultural products. Ministry of Agriculture and Forestry 3-5. Lee KH, Kang SJ, Choi BD, Choi YJ and Youm MG. 1994. Utilization of ascidian tunic extracts on pigmentation and growth of rainbow trout. J Fish Sci Tech 27, 232-239. Park HY, Lim CW, Kim YK, Choi TJ, Yoon HD, Lee KJ, Seo YK, Kim JY and Park KE. 2007. Effects of supplemental asterias amurensis extract in the experimental diets on growth, blood chemistry and superoxide production of kidney phagocytes of Sebastes schlegeli. J Korean Soc Appl Biol Chem 50, 362-366. Schut HA, Cumming DA and Smale MHE. 1997. DNA Adduts of heterocyclic amines: Formation, removal and inhibition by dietary components. Mutat Res Fund Mol Microbial 376, 185-194. Tee ES. 1992. Carotenoids and retinoids in human nutrition. Cri Rev Food Sci Nutr 31, 103-163 Torrissen OJ. 1986. Pigmentation of salmonids A comparison of astaxanthin and canthaxanthin as pigment sources for rainbow trout. Aquaculture 53, 271-278. Twibell RG, Watkins BA and Brown PB. 2001. Dietary conjugated linoleic acids and lipid source alter fatty acid composition of juvenile yellow perch, Perca flavescens. J Nutri 131, 2322-2328. Umeda K, Tanka Y and Ohira O. 1971. Carotenoid pattern of citrus unshiu flesh analysis of orange juice. Nippon Shokuhiu Kogyo Gakkaishi 18, 13-20. Whang HJ. 1999. The change of carotenoid pigment in Korean pumpkin using drying. Food Eng Prog 3, 214-219. 2010 년 8 월 20 일접수 2010 년 10 월 12 일수정 2010 년 12 월 3 일수리