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Journal of Life Science 2015 Vol. 25. No. 9. 1072~1079 Application of the Lees of Domestic Traditional Wine and its Useful Biological Activity Mi-Sun Kim 1, Woo-Chang Shin 2 and Ho-Yong Sohn 1 * 1 Department of Food and Nutrition, Andong National University, Andong 760-749, Korea 2 Research Institute, Kooksoondang Brewery Co. LTD., Seongnam 460-120, Korea Received September 18, 2015 /Revised September 29, 2015 /Accepted September 30, 2015 The lees of Korean traditional wine called as Jubak or Sul-jigemi in Korea is byproduct from alcohol fermentation industry, which is remnant of fermentation broth after filtration, centrifugation, distillation, or sedimentation during aging. Since, Korean traditional wines are produced from edible plant sources such as rice, foxtail millet, fruits and medicinal herbs with nuruk (a traditional fermentation starter and starch degrader), the Jubak from rice wine (takju), medicinal herb wine (yakju) and fruits wine are considered as safe byproduct and have various useful bioactivity. Considering the recent rapid increased production of Jubak from takju industry, and the reinforcement of dispose of Jubak as waste material in worldwide, the development of efficient reuse process for Jubak is necessary in traditional wine industry. In this review, the status of current industry, research and patent trends in relation with Jubak production, treatment, utilization and renewal was analyzed and different bioactive compounds including phenolic acids from Jubak were provided. Jubak is not any more waste material, and is the source of bioactive functional materials for food, cosmetics and medicinal industry. To develop the efficient and economic renewal technology including recovery process for bioactive substances from Jubak, systematic collaboration and research among the industry, academy and government is necessary. Key words : Bioactivity, bioactive substances, jubak, lees of Korean traditional wine ISSN (Print) 1225-9918 ISSN (Online) 2287-3406 DOI : http://dx.doi.org/10.5352/jls.2015.25.9.1072 - Review - 서 알코올은탄화수소의수소원자가하이드록시기 (-OH) 로치 환된화합물의총칭으로분자량에따라상온에서액체또는 고체로존재하며, 하이드록시기의치환수에따라 1 가알코올 ( 에탄올 ), 2 가알코올 ( 에틸렌글리콜 ), 3 가알코올 ( 글리세린 ) 등 으로구분가능하다. 알코올류중메탄올, 에탄올, 부탄올등의 저분자알코올은인체에독성을나타내어저독성 (slightly toxic: 경구치사량 5~15 g/kg) 이상으로규정되어있다. 한편술은 알코올성분이들어있어마시면사람이취하게되는음료의 총칭으로, 우리나라주세법에서는알코올함량 1 도이상의음 료를말한다 [25]. 따라서술에서알코올은구체적으로에탄올 을의미하고있다. 술은인류가만든최초의발효음료로, 발효 중상당량의이산화탄소가발생하여, 마치물 ( 수 ) 속의끓는불 처럼보여수불로불렸으며, 이후수울, 수을에서다시술로 변화하였다고알려져있다 [25]. 따라서술은물속의불이라는 *Corresponding author *Tel : +82-54-820-5491, Fax : +82-54-820-7804 *E-mail : hysohn@anu.ac.kr This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 론 신성함과정신이아찔해지는혼돈성을동시에가지고있어, 단순한음료이상의의미를가지고있으며신과인간과의소통및인간과인간의소통에이용되고있어인류가존재하는한알코올산업은지속될것이다. 현재국내알코올산업은주류용, 산업용및기타주정소비분야로구별할수있으며, 연간 8.6조원의시장을형성하고있다. 최근에는외국주정수입및시장자율화, 외국와인및맥주류의수입증대, 전통주의소비감소등으로인해전분야에서어려움을겪고있으나지속적인발효균주육종, 액화당화효소개발, 원료전분의효율적이용, 증류공정개선, 다양한전통주발굴등으로극복해나가고있다. 그러나발효폐기물관리측면에서는아직개선이이루어지지않고있는실정이다. 발효폐기물의대부분을차지하는주박은발효후알코올성발효액을필터, 원심분리또는증류후에얻어지는찌꺼기를말하며, 보다광의의주박은 1차제조된제품의숙성과정중에생성되는찌꺼기까지포함한다 [33]. 주박은발효시의사용원료 ( 발효기질 ), 사용누룩, 발효방법및제성방법에따라그성분이나영양적특성이다양하게변화될수있으며 [16, 17], 주로미생물균체, 미분해원료기질, 발효생산물, 유기산, 미네랄등을함유하고있어높은 BOD 및 COD를나타내어산업폐기물로관리되고있는실정이다. 특히아레기로불리는증류후폐액은 ph가낮으면서도환원성이강해토양비료용및가축사료용으로도부적합상태이며, 전통주발효에서사용

Journal of Life Science 2015, Vol. 25. No. 9 1073 되는백미, 찹쌀등의발효기질과는달리, 산업용주정생산의발효기질들은타피오카, 카사바, 고구마전분등이주를이루어잠재적유해물질 ( 잔류농약, 중금속등 ) 의문제로그이용이매우제한되는실정이다. 따라서주류산업에서의가장큰고민은 발효부산물 ( 주박 ) 을어떻게처리할것인가? 로, 이의배경에는 2006년 3월산업폐기물해양투기금지협약의발효, 2012년가축분뇨, 하수슬러지해양투기금지, 2013년음식물쓰레기및발효주박해양투기금지및 2016년육상폐기물해양투기전면금지의법적규제가관련되어있다. 따라서주박의효율적인처리는향후알코올및전통주산업의지속발전에가장중요한문제가되리라예상된다. 전통주발효주박의우수성우리나라는 1월의귀밝이술에서부터 5월단오의창포주, 8 월추석의햅쌀술, 11월동지의동정춘등세시풍속과밀접한관련이있는수많은술을만들어음용하여왔으며, 유교문화중심의제례를중시하던조선시대에는 600 여종이상의가양주가제조되었다고알려져있다 [25]. 이러한전통주들은현대의산업용주정제조와는달리, 다양한식용및약용식물원재료를발효기질로사용하며, 산업용액화, 당화효소보다는누룩을사용하며, 지역마다다양한발효효모를사용하므로, 부산물인주박또한다양한특성을나타낸다 [20, 25]. 특히발효시에당화제및종균제의역할을동시에수행하는누룩은우리나라에서만사용하는독특한발효개시제로통상, 통밀, 백미, 보리, 조등의식용재료를이용하여제조되며, 지역마다누룩에사용하는재료와제조방법이다양하다 [19, 23, 36]. 이에따라, 사용누룩에차이에따른우점종균주들의다양성이나타나며, 같은술이라하더라도지역마다제조된술의관능성및유용생리활성에많은차이가나타나게된다. 이는발효부산물인주박에서도다양한기능성을나타내게하는이유이기도하다. 또한전통주로부터발생하는주박은산업용주정제조시발생되는주박과는달리식용재료, 한방약재, 고품질누룩을사용하여안전성을확보하고있다 [15, 16, 17]. 따라서전통주주박은산업폐기물이라기보다는유용기능성에따른고부가가치제품개발원료로사용가능한미생물발효전환산물원료라는인식의변화가필요하다. 국내전통주주박의산업동향국내의전통주산업은전체주류시장 ( 약 8.6조원 / 년 ) 의 8% 를차지하는것으로알려져있으며, 산업규모는 6,900억원에이르고있다. 최근정부의막걸리산업지원과함께사회적웰빙분위기에따라탁주 ( 막걸리 ) 시장은빠르게성장해왔다. 실제 2011년을정점으로, 탁주시장의성장세가다소주춤한상태이나, 탁주는우리나라의대중주로확고히자리매김하고있는만큼탁주주박의생산은지속될것으로예상된다 (Table 1). 최근의전통주는백미를주재료로하고있으나, 다양한약 Table 1. Changes on delivery of domestic traditional wine from a warehouse (Unit: Kl) Year Takju Yakju Fruit wine Sum 2006 2007 2008 2009 2010 2011 2012 2013 170,165 172,370 176,398 260,701 412,279 458,198 448,047 426,216 42,873 33,288 27,374 22,116 18,841 18,975 16,544 14,538 45,046 61,127 56,015 45,972 46,915 46,384 46,505 52,091 * From National Tax Service (2014. 12. 19) 258,084 266,785 259,787 328,789 478,035 523,557 511,096 492,845 용식물, 한약재를포함하는다양한기능성약용주가주로개발되고있다. 또한지구온난화에따른한반도기온증가에따라고도주보다는저도주가인기가있으며, 전통주는소주, 양주와달리낮은도수와건강지향성으로시장이지속적으로증가되리라예상된다 (Table 1). 한편전통주주박과관련하여, 정확한발생량은알수없으나, 연간 16,000톤이상의주박이발생하는것으로추정되고있으며, 이의 60% 이상은탁주주박으로판단된다 (Table 2). 통상탁주의경우주박발생비율은원재료의 2.3%, 약주의경우 15% 전후이며, 복분자주와같은과실주의경우 7.5% 전후로알려져있다. 따라서, 상대적으로탁주주박발생비율이약주나과실주에비해 15~30% 수준으로낮은상태이나, 국내주박발생의 60% 이상이탁주주박인것은, 탁주의생산량이월등히높아이에따른주박발생량도증가하기때문이다 [17]. 전통주주박의경우사용원료 ( 발효기질 ), 사용누룩, 발효방법및제성방법에따라그특성이변화되나, 통상, 탁주주박은일반적인제상방식으로생산되며 ph 3.9~4.5, 수분함량은 60~ 80%, 알코올함량은 10~16%, 잔당함량은 3~4% 를포함하는뻑뻑한액체형태이며, 약주주박의경우압착방식을사용하여생산되며, ph 3.7~4.3, 수분함량은 35~50%, 알코올함량은 5~7%, 잔당함량은 4~5% 를포함하는유통성고체형태이다 [16]. 따라서, 전통주주박은높은 BOC, COD를나타내며, 현재대부분의전통주제조회사에서는주박을사료로일부이용하거나, 폐기물처리업체에위탁처리하게된다. 반면맥주박 Table 2. Estimated annual production of lees of traditional wine (Unit: ton) Year Takju Yakju Fruit wine Sum 2006 2007 2008 2009 2010 2011 2012 2013 3,914 3,965 4,057 5,996 9,482 10,539 10,305 9,803 6,517 5,060 4,161 3,362 2,864 2,884 2,515 2,210 3,469 4,707 4,313 3,540 3,612 3,572 3,581 4,011 13,899 13,731 12,531 12,898 15,959 16,994 16,401 16,024

1074 생명과학회지 2015, Vol. 25. No. 9 또는비전통주주박은효모추출물제조, 사료및퇴비로이용되고있다. 따라서증가되고있는주박생산량을고려한다면, 이의효율적인처리기술과함께고부가가치화산업구조확립이필요한실정이다. 국내전통주주박관련연구동향 1990년부터현재까지국내에보고된주박관련연구는 Table 1에나타내었다. 2000년이전에는주로주박을이용한장아찌제조, 주박의처리에의한사료및퇴비화연구 [12, 23], 다양한곡류가공식품에첨가하는식품첨가물로연구가주로진행되었으며, 최근에는식품부재료, 미생물발효배지조성물 [28, 29], 고부가가치기능성식품소재로활용하기위한기능성탐색등이주로보고되고있다. 현재까지가장많이연구된분야는주박의유용생리활성에대한연구 (48%) 와식품첨 A B C s Fig. 1. Relative frequency of research fields for the lees of traditional wine (A), the reported useful biological activity of the lees of wine (B) and the usage of less of wine as food additives (C).

Journal of Life Science 2015, Vol. 25. No. 9 1075 Table 3. Current domestic researches in relation with less of Korean traditional wine No. Research title Lees origin Reference 1 2 3 4 Anti-microbial, anti-oxidant, and anti-thrombosis activities of the Lees of Bokbunja Wine (Rubus coreanus Miquel) Evaluation of in-vitro antithrombosis and anti-oxidation activity of lees of takju (Wookukseng) Anti-adipogenic, anti-inflammatory, and anti-proliferative activities of extracts from Lees and Nuruk Anti-proliferative and Anti-inflammatory Activities of Solvent Fractions of Lees Extracts Fruits and Yakju 5 Quality Characteristics of Muffin added with Lees 6 7 8 9 10 11 12 13 Anti-inflammatory Effects of Extracts and Their Solvent Fractions of Rice Wine Lees Suppressive Effects of Extracts and Solvent Fractions of Rice Wine Lees on Inflammation in RAW 264.7 cells Evaluation of in-vitro Antithrombosis Activity of Lees of Korean Traditional Wine Skin Whitening and Anti-Wrinkle Effects of Extract from Jubak of Oriental Herbal Liquor Anti-proliferative Activities of Solvent Fractions of Lees Extracts in Human Colorectal HCT116 Cells Anti-microbial and Anti-thrombosis Activities of Lees of Sweet Potato Soju Quality Characteristics and Antioxidant Activity of Jeung-pyun added with Ju-bak Powder Anti-obesity, Anti-oxidation, Anti-inflammation and Anti-cancer Activities of Jubak and Nuruk Extracts 14 Quality Characteristics of Sulgidduk with Lees 15 16 17 18 19 20 21 22 23 Antioxidant Activity and Quality Characteristics of Rice Wine Cakes Cookies with Different Ratio of Astragalus memvranaceus Quality characteristics of cucumber Jangachi using Sake cake by Salt concentration and storage period Inhibitory Effect on Tyrosinase, ACE and Xanthine Oxidase, and Nitrite Scavenging Activities of Jubak (Alcohol filter cake) Extracts Quality Characteristics of Low-Salt Yacon Jangachi Using Rice Wine Lees during Storage Ethyl Acetate Extract of Korean Rice Wine Lees Inhibits IgE-Mediated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells and Passive Cutaneous Anaphylaxis in Mice Dietary Fiber from Lees of Unpolished Rice : Clinical Effect on the Obese Anti-Wrinkle Effects of Korean Rice Wine Cake on Human Fibroblast Antimicrobial, Antioxidant and SOD-Like Activity Effect of Jubak Extracts Effects of lees from brown rice on waist circumference and Metabolic Parameters in Type 2 Diabetes: A Randomized, Double-blinded, Placebo-controlled Trial Kim MS et al., 2015, [14] Kim MS et al., 2015, [17] Son JB et al., 2015, [35] Kang HT, 2015, [7] Yun CS et al., 2014, [38] Park MJ et al., 2014, [31] Park MJ, 2014, [30] Kim MS et al., 2014, [16] Lee SM et al., 2014, [26] Kang HT et al., 2014, [8] Kim MS et al., 2014, [15] Ko YS & Shim KH, 2014, [21] Son JB. 2013, [34], Root of Astragalus memvranaceus Cho YH et al., 2013, [3] Lim, JM et al., 2013, [27] Chung-ju Park, YR. 2013, [32] Rice wine Kwon SC et al., 2012, [22] Jung, HN et al., 2012, [5] Kang YJ et al., 2011, [9] Dietary fiber Lee MS, 2011, [25] Yoo JM, 2010, [37] Kim TY, 2010, [20] Fermented Dietary fiber from Kim EK, 2010, [10]

1076 생명과학회지 2015, Vol. 25. No. 9 Table 3. Continued No. Research title Lees origin Reference 24 Research of Effective Method for Pellet Using Jubak 25 26 27 28 29 30 Effects of Distiller Solubles (Bekseju) Application on the Growth of Rice Plant (Oryza sativa L.) and Improvement of Soil Fertility Qualitative Characteristics of Brown Sauce with added Jubak and Chicken Meat marinated in Jubak Isolation and characterization of tartaric acid-degrading bacteria from Korean grape wine pomace Characteristics of Ju-Back and Effect of Ju-Back Fertilizer on Growth of Crop Plants Physicochemical Characteristics of Yackwa with Added Rice Wine Cake Quality Characteristics of Noodle added with Takju (Korean turbid rice wine) lees,yak ju, Chung-ju Yakju Kim DN et al., 2010, [11] Kim IS et al., 2009, [12] Lee JP, 2008, [24] grape wine pomace 31 Quality Characteristics of Pork with Addition of Jubak (Sulchigegie) 32 33 Effects of Takju (Korean turbid rice wine) Lees on the Serum Glucose levels in Streptozotocin-induced Diabetic Rats Effects on Quality Characteristics of Pork Loin Fed with Wild Grape (Vitis amurensis Ruprecht) Wine By-product Fruit wine (Vitis amurensis) 34 Production of Yeast Spores from Rice Wine Cake 35 36 Effects of Spray Dried Takju Cake Powder on the Quality of White Pan Bread Selection of optimum medium for mass production of Metarhizium anisopliae ARS 978 37 Edible Films from Protein Concentrates of Rice Wine Meal Kim, JH et al., 2008, [13] Lee JH et al, 2007, [23] Cho EJ et al. 2007, [1] Kim SM et al. 2007, [19] Won JH et al. 2006, [36] Kim SM et al. 2006, [18] Jung IC et al. 2005, [6] Lim YS et al., 2004, [28] Jung BJ. 2002, [4] Protein from Moon KH et al., 1998, [29] Cho SY et al., 1998, [2] 가물이용분야 (32%) 이며, 비료및사료개발, 발효배지로이용, 식용필름제조및바이오에너지생산분야도보고되어있다 (Fig. 1A). 한편, 기존의보고된주박의생리활성으로는항산화활성 [14, 17, 20, 21, 27] 이가장많으며, 최근에는항혈전 [14, 15, 16, 17], 항염증 [7, 30, 31, 34, 38], 항균 [14, 15, 20] 및암세포성장억제활성 [7, 8, 38] 들이보고되고있다. 그외, 항주름및미백활성 [26, 37], 항당뇨 [10, 18], 알러지감소 [9], 항비만및고혈압억제활성 [22, 25, 34] 도알려져있어 (Fig. 1B), 전통주주박은다양한생리활성을가지고있음이확인되어있다. 전통주주박의식품부재료로서의적용성연구로는떡류 [3, 21], 장아찌 [5, 32], 쿠키 [27], 돼지고기가공 [6, 36] 등에집중되어있으며, 그외머핀 [38], 소스 [24], 약과 [1], 국수 [19], 케이크제조 [4] 등에도사용되고있다 (Fig. 1C). 향후, 기능성과안정성이확보된전통주주박을이용한식품소재화연구는빠르게진행될것으로예상된다. 국내전통주주박관련특허동향국내의주박관련특허는현재 57건이알려져있으며, 그 중등록특허는 15건이다. 등록특허들은식품첨가물로주박이용성에관한특허가 7건으로전체의 47% 를차지하며, 이어 4건의기능성향장및화장품소재로의이용, 3건의기능성식품소재로이용및 1건의바이오에너지생산관련이알려져있다 (Table 3). 반면등록을기다리고있는주박특허출원의경우식품첨가물관련이 11건으로가장많으나, 기능성식품소재관련8 건, 기능성향장제품관련 3건, 사료이용 4건, 비료이용 2건, 에너지화 2건으로, 최근에는주박의신규유용기능성관련특허가증가되고있는실정이다. 향후에는이러한주박의기능성발굴과함께, 다양한제품이생산된다면, [ 주박 = 폐기물 ] 이라는인식이 [ 주박 = 중요발효기능성소재공급원 ] 으로전환되리라예측된다. 국내전통주주박의유용생리활성물질일본및유럽의경우일본주및와인으로부터 kojic acid, ferulic acid 및 resveratrol과같은다양한유용생리활성물질들이보고되어있다 [18]. 한국의전통주에서는 farnesol 및 squalene이보고된바있으나, 보다많은활성물질의분리확

Journal of Life Science 2015, Vol. 25. No. 9 1077 Table 4. Current domestic patent in relation with less of Korean traditional wine No Patent No. Title of invention 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 1012701180000 1010575980000 1014060980000 1013833740000 1011526360000 1012101490000 1014510960000 1014507730000 1011194140000 1013974050000 1014305910000 1012396300000 1012709170000 1014355660000 1014030320000 Low-Fat Chicken sauce containing makgeolli lees fiber and manufacturing methods thereof Rice fermented bread and method of producing the same Chicken meat patties comprising brewers grain dietary fiber and the manufacturing methods Rice muffin that contain lees and pocedd of that muffin Jangajji and method for preparing the same Manufacturing method of dombaeki by process of residue left after rice liquor is drained Preparing method of low-salt umeboshi using Rubus coreanus jubak (Alcohol filter cake) Glycoprotein fraction isolated from rice wine lees and method for producing the same Composition for enhancement of immune function which comprises rice wine lees as an active ingredient Preparing method of concentrated extract of Rubus coreanus using Rubus coreanus jubak (Alcohol filter cake) Fermented cosmetics Soap and methods for manufacturing methods Functional bath cleaner composition Compound and method on power containing maggulri ingredients Method for preparing ethanol using residual product of brewery A B C D E F G H J I Fig. 2. Biologically active phenolic compounds in the less of Korean traditional wines. (A) gentisic acid, (B) chlorogenic acid, (C) caffeic acid, (D) cinnamic acid, (E) syringic acid, (F) coumaric acid, (G) ferulic acid, (H) gallic acid (I) vallinic acid, and (J) sinapic acid. 인이필요한실정이다. 최근다양한탁주, 약주및과실주 ( 복분자주 ) 의전통주주박의에탄올추출물로부터항산화, 항혈전활성을나타내는다양한페놀성화합물 (phenolic compound) 을분리하였으며, 이들은 gentisic acid, chlorogenic acid, caffeic acid, cinnamic acid, syringic acid, coumaric acid, ferulic acid, gallic acid, vanillic acid 및 sinapic acid로확인되었다 (Fig. 2). 이러한물질들은이미항산화, 항균, 항혈전, 항염증, 암세포성장억제, 미백및주름개선활성등이보고되어있어전통주주박의유용생리활성의주요활성물질로추측된다. 향후상기분리된물질들의각각의유용생리활성관련메커니 즘구명이필요하며, 전통주주박으로부터이러한활성물질을효율적으로회수할수있는추출, 정제방법의연구도필요한시기이다. 결론전통주의주박은식용백미를주로사용하면서다양한약용식물이첨가되고, 이를누룩으로발효시켜안정성이우수하면서, 또한다양한유용생리활성들을가지는것으로알려져있다. 그러나환경에그대로방출시에는다양한환경오염문제를

1078 생명과학회지 2015, Vol. 25. No. 9 야기하므로, 발효폐기물로지정되어있는실정이다. 주박의효율적인재처리는, 전통주탁주시장의성장으로주박생산량이빠르게증가되고있는현재, 가장중요한문제가되고있다. 따라서전통주주박의효율적인재처리를통해주박을식량 ( 식품재료 ), 사료, 퇴비, 미생물 ( 효모및유산균 ) 배양원및다양한산업용추출물 ( 효모추출물등 ) 로개발하여야하며, 나아가전통주주박의유용생리활성발굴및유용물질확인을통해, 기능성식품, 의약품, 향장소재개발의중요발효자원으로활용하여야한다. 최근탁주, 약주, 및과실주의전통주주박으로부터확인된다양한 phenolic acid 화합물들의유용생리활성기작연구와이들의대량분리, 회수연구가필요하며, 이를위한전통주주박과관련된체계적인산학관연연계와연구가필요하다. 감사의글 본연구는 2012년도농림수산식품부고부가가치식품기술개발사업 ( 과제번호 112073-3) 에의해수행되었으며, 이에감사드립니다. References 1. Cho, E. J., Yang, M. O. and Kang, H. J. 2007. Physicochemical characteristics of Yackwa with added rice wine cake. J. East Asian Soc. Dietary Life 17, 94-102. 2. Cho, S. Y., Park, J. W. and Rhee, C. 1998. Edible films from protein concentrates of rice wine meal. Kor. J. Food Sci. Technol. 30, 1097-1106. 3. Cho, Y. H., Cho, J. S., Kim, J. Y., Kim, U. S., Choi, J. H. and Park, J. H. 2013. Quality characteristics of sulgidduk with makgeolli lees. J. East Asian Soc. Dietary Life 23, 227-233. 4. Jung, B. J. 2002. 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