Journal of Radiation Industry 7 (1) : 29~35 (2013) Review Paper 감마선조사를이용한시판분말원료의위생화 최수정 한인준 윤영민 김종헌 김재훈 김재경 박종흠이주운 홍성길 1 육홍선 2 송범석 * 한국원자력연구원첨단방사선연구소 1 ( 주 ) 이롬생명과학연구원, 2 충남대학교식품영양학과 Sanitization of Commercial Powdered Products Using Gamma Irradiation Soo-Jeong Choi, In-Jun Han, Young-Min Yoon, Jong-Heon Kim, Jae-Hun Kim, Jae-Kyung Kim, Jong-Heum Park, Ju-Woon Lee, Seong-Gil Hong 1, Hong-Sun Yook 2 and Beom-Seok Song* Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup 580-185, Korea 1 Erom R&D Center, Gangwon 200-944, Korea 2 Department of Food and Nutrition, Chungnam National University, Daejeon 305-764, Korea Abstract - Microbiological populations and the sterility of commercial powdered products treated with gamma irradiation at 0~10 kgy were investigated before using them as ingredients for a non-cooked Saengsik product. We evaluated a total of 14 powdered products: 8 powdered cereals, 3 powdered tubers, and 3 powdered leafy vegetables. The total numbers of bacterial populations in non-irradiated powdered cereals, tubers, and leafy vegetables were 2.7~6.9, 5.6~6.0, and 5.3 ~6.8 log CFU g - 1, respectively. Moreover, coliform bacteria were not indicated in adlay, millet, germinated brown rice, soybean, and mulberry leaves powder within detection limit (2.0 log CFU g - 1 ). The number of Bacillus cereus exceeded 3.0 log CFU g - 1 (the maximum limit for Saengsik products) in all samples, excluding perilla seeds, buckwheat, barley, oat, potato, and Jerusalem artichoke powder. However, a dose of 6 kgy of gamma irradiation reduced the microbiological populations in all samples, and all the powdered products met the microbial requirements for Saengsik products. Futhermore, it was confirmed that all microorganisms in the 9 powdered products, except fermented brown rice, sweet potatoes, and 3 leafy vegetables, were sterilized by 10 kgy of gamma irradiation. Key words : Powder, Gamma irradiation, Saengsik, Microbiological population, Sterilization * Corresponding author: Beom-Seok Song, Tel. +82-63-570-3211, Fax. +63-570-3207, E-mail. sbs0110@kaeri.re.kr 29
30 최수정 한인준 윤영민 김종헌 김재훈 김재경 박종흠 이주운 홍성길 육홍선 송범석 서론국민소득증가와평균수명이증가함에따라삶의질이향상되었으며, 이에따른건강에대한관심도가높아져안전하면서건강에도움이되는건강요구형식품의관심이증가하고있다 (Chang et al. 2004). 그에따라건강기능성식품의수요가급증하고있는데, 그중에서도생식제품이주목을받고있다. 생식은동 식물성원료를주원료로건조등가공처리하여분말, 과립, 바, 페이스트, 겔, 액상등으로제조한것으로이를그대로또는물등과혼합하여섭취할수있도록편리성을지니게한것을말한다 (KFDA 2012). 생식의국내시장규모는 2000년 900억원에서 2002년 2,000억원, 2005년에는약 5,000억원수준으로증가하였으며, 특히 1998년이후부터미국, 중국및일본등총 10여개국가에대한수출액이약 1,500만달러로성장하였다 (Lee 2004). 생식제품의원료들로는곡류, 채소류, 버섯류, 해조류및과일류등이사용되고있으며생식의주요한원료인채소류, 버섯류및해조류에는 3대영양소, 미량영양소, 식이섬유소등을포함한다양한기능성물질들을다량함유하고있다 (Lee et al. 2003; Lee et al. 2003; Park et al. 2003). 이러한기능성물질들은콜레스테롤저하및변비억제등의전통적생리활성을벗어나서최근에는항암작용및항균작용등다양한생물활성을나타내는것으로보고되고있다 (Cho et al. 2008). 분말원료는대부분신선한채소, 과일및곡류를사용하기때문에수확후세척, 정선또는탈피, 건조과정을거쳐제조된다 (Lee 2002; Kim et al. 2004). 따라서이러한비가열공정으로인해원료내에존재하는미생물또한그대로제품에존재하거나증식하는문제점을내포하고있다 (Chang et al. 2004). Chang et al. (2004) 은일부시판생식제품에서식중독균인 Bacillus cereus와대장균군이검출되어이들에대한미생물학적안전성에대한문제를제기한바있다. 미생물오염은원료의품질저하뿐만아니라섭취시에도조리를하지않고단순음용하는과정에서위생학적문제를야기시킬수있으므로, 이를개선시키기위한위생화공정이필요하다 (Kim et al. 2002). 한편방사선조사기술은대표적인비가열살균기술로서식품고유의풍미와생화학적품질, 그리고기능성을유지하면서도제품을포장한상태에서조사처리가가능해 2차오염을방지할수있다 (Thayer 1990; Byun and Lee 2003; Lee et al. 2003). 이러한방사선조사기술은살 균또는멸균이필요한건조식품, 냉동식품및분말식품등에응용이가능하여우주식품, 전투식량및환자식으로도활용되고있다 ( 김등2009). 이와같은특성을가지는방사선조사기술로생식과같이가열살균이어렵고기능성을유지해야하는제품의살균이가능하다 (Kim et al. 2002). 따라서본연구에서는생식제품의원료로사용가능한시판분말제품들의미생물오염수준을확인하고, 이들을살균또는멸균할목적으로방사선조사기술을적용한후미생물사멸정도와멸균선량을확인하고자하였다. 재료및방법 1. 재료본실험에사용된분말원료는곡류 ( 율무, 들깨, 기장, 메밀, 발아현미, 보리, 백태, 귀리 ), 서류 ( 감자, 돼지감자, 고구마 ), 엽채류 ( 샐러리, 케일, 뽕잎 ) 분말로인터넷을통해구매하였으며, 미생물오염도를조사하기위해 Difco Laboratories (Detroit, Michigan, USA), Oxoid (Hampshire, England), 3M (St. Paul, MN, USA) 에서배지를구입하여사용하였다. 2. 감마선조사시판분말제품의감마선조사는선원 11.1 PBq, 60 Co 감마선조사시설 (IR-70 gamma irradiator, MDS Nordion, Canada) 을이용하였다. 샘플 50 g을각각ldpe (low density polyethylene) 에포장한후각각 0, 2, 4, 6, 8 및 10 kgy 의흡수선량 ( 선량률 10 kgy h -1 ) 이되도록조사하였다. 조사후총흡수선량확인은 alanine dosimeter (Ceric cerous dosimeter, Bruker Instruments, Rheinstetten, Germany) 를사용하였다. Dosimetry 시스템 (Kishor et al. 1995) 은국제원자력기구 (IAEA) 의규격에준용하여표준화한후사용하였으며, 총흡수선량의오차는 2% 이내였다. 3. 미생물검사시료의일반호기성미생물, 진균류, Bacillus cereus, Clostridium perfringens 및대장균군의미생물생육정도를확인하였다. 즉, 시료 10 g에멸균된 peptone수 90 g을가하여 1분간 stomacher (Mark II Lab Blender, Tekmar Teledyne Technologies Inc., Sacramento, CA, USA) 를이용하여균질화한후 10진희석법으로희석한희석액을실험에사용하였다. 일반호기성미생물은 plate count agar (PCA,
시판분말원료의감마선위생화 31 Difco Co. Detroit, MI, USA), 곰팡이는 potato dextrose agar (PDA, Difco Co. Detroit, MI, USA), B. cereus는 mannitol egg yolk polymyxin agar (MYP, Difco Co. Detroit, MI, USA), Cl. perfringens는 tryptose sulfite cycloserine agar (TSC, Oxoid, Hampshire, England) 의 plate에각각도말하였고, 대장균군은 3M 건조필름배지 (3M Petrifilm, St. Paul, MN, USA) 에접종하였다. 도말된 PCA, MYP 배지는 35 C 에서 2일간, TSC는 anaerobic jar (Oxoid, Hampshire, England) 를이용하여혐기적조건에서 35 C 에서 2일간, PDA 배지는 25 C 에서 3~5일간각각배양하여 30~300개의집락을형성한배지만을계수하였다. 접종한건조필름배지는 35 C 에서 1일간배양하여생성된붉은집락중주위에기포를형성한집락수를계산하였다. 4. 세균발육시험시료의멸균여부를확인하기위해식품공전의세균발육시험법 (KFDA 2012) 을이용하였다. 즉, 시료를무균적으로취한후멸균 PBS buffer solution 45 ml를가하고, 20초간 vortexing 하였다. 이용액의 1.0 ml에멸균된 thioglycollate medium (Sigma-Aldrich co., St. Louis, MO, USA) 을가하고, 35 C 의 incubator (MIR-262, Sanyo, Tokyo, Japan) 에서 48시간동안배양시켜 yellow 계통으로변색되지않는것을멸균된것으로확정하였다. 결과및논의 1. 시판분말제품중곡류분말의미생물평가시판분말제품중곡류분말 8종 ( 율무, 들깨, 기장, 메밀, 발아현미, 보리, 백태및귀리 ) 의미생물오염수준과방사선조사에따른미생물변화및멸균여부확인결과를 Table 1에나타내었다. 율무, 들깨, 기장, 메밀, 발아현미, 보리, 백태및귀리의일반호기성미생물은비조사구에서 2.7~6.9 log CFU g -1 으로나타났으며, 특히율무, 들깨및발아현미분말은 5.0 log CFU g -1 이상으로높게나타났다. 이는 Kim et al. (2012) 의현미와보리분말에서일반호기성미생물의초기생육도가 3.0~5.0 log CFU g -1 으로나타났다는결과와유사하였다. 또한비조사구의진균류는 2.4~3.5 log CFU g -1 이였고대장균군의경우음성반응으로검출되지않은들깨를제외한 7종의곡류분말에서양성반응으로나타났고, Cl. perfringens는곡류분말8종모두에서검출되지않았다. 그러나 B. cereus의경우검출되지않은들깨와귀리및 3.0 log CFU g -1 이하로나타난메밀, 보리를제외한나 머지분말원료들에서는 3.0 log CFU g -1 이상으로나타나식품공전에서제시하고있는생식제품에대한미생물기준 (Table 4) 에부적합한것으로판단되므로들깨, 메밀, 보리, 귀리분말을제외한나머지 4종의곡류분말은생식제품원료로사용이불가능할것으로사료되었다. 감마선조사처리에의하여모든미생물수는감소하였으며, 곡류분말 8종모두6kGy 이하의선량으로감마선조사했을때식품공전의생식제품미생물기준을만족하는것으로나타났다. 세균발육시험을통한감마선조사된곡류생식분말의멸균확인실험결과에서들깨, 기장, 보리및귀리는 4 kgy, 메밀은 6 kgy, 율무및백태는 8kGy에서음성반응이나타내어분말제품각각의멸균선량을확인할수있었다. 그러나발아현미의경우는초기균수가 6.3 log CFU g -1 으로매우높아 10 kgy에서도멸균되지않은것으로나타났다. 2. 시판분말제품중서류분말의미생물평가시판분말제품중서류분말 3종 ( 감자, 돼지감자및고구마 ) 의미생물오염수준과감마선조사처리된시료의일반호기성미생물, 진균류, B. cereus, 대장균군및 Cl. perfringens의생육정도및멸균여부확인결과는 Table 2에나타내었다. 감자, 돼지감자및고구마의일반호기성미생물은비조사구에서각각 5.8, 5.6 및 6.0 log CFU g -1 으로나타났다. 이는감자의초기일반호기성세균이 5.0 log CFU g -1 의수준으로나타났다는 Kim et al. (2012) 과 Koh and Kim (2010) 의결과와유사하였다. 비조사구의진균류는감자, 돼지감자및고구마각각 2.3, 2.8 및 2.7 log CFU g -1 으로서로유사한수준을보였고대장균군은각각 2.5, 1.9 및 2.8 log CFU g -1 으로검출되었으며, Cl. perfringens는서류분말3종모두에서검출되지않았다 ( 검출한계 10 CFU g -1 ). B. cereus의경우고구마분말에서 5.4 log CFU g -1 으로생식제품미생물기준 (3.0 log CFU g -1 ) 을초과하여분말자체를이용한생식제품으로의제조는불가능한것으로판단되었다. 서류분말 3종의경우모두 4 kgy 이하의선량으로감마선조사했을때생식제품의미생물기준 ( 식품공전 2012) 을충족하였다. 감마선조사된서류생식분말의멸균확인실험결과에서감자 8 kgy 및돼지감자 10 kgy 에서멸균이확인되었으나고구마는 10 kgy에서도멸균되지않은것으로나타났다. 이것은서류작물은토양에서자라는덩이뿌리식물로서토양미생물인 B. cereus외에도다른다양한미생물군이제조공정에서충분히제
32 최수정 한인준 윤영민 김종헌 김재훈 김재경 박종흠 이주운 홍성길 육홍선 송범석 어가되지않을경우많은수의세균들이최종분말제품에존재할가능성이매우높으므로 (Cho et al. 2008), 높은초기오염도로인하여멸균선량이상승한것으로사료되었다. Table 1. Effect of gamma irradiation on bacterial populations in powdered cereals and bacterial growth after incubation 35 C for 48 h Samples Adlay Perilla seeds Millet Buckwheat Microbes (log CFU g -1 ) Irradiation dose (kgy) Total aerobic Bacillus Clostridium Coliform Bacterial Molds bacteria cereus perfringens bacteria growth 0 5.3±0.03 3.3±0.05 5.4±0.01 ND TNTC 2) + 3) 2 3.8±0.16 2.3±0.28 3.5±0.06 ND ND + 4 3.1±0.35 ND 2.5±0.07 ND ND + 6 0.7±1.15 ND 1.3±1.15 ND ND + 8 ND 1) ND ND ND ND - 0 5.0±0.18 2.7±0.13 ND ND ND + 2 2.3±0.35 ND ND ND ND + 0 4.7±0.09 3.4±0.08 3.5±0.03 ND TNTC + 2 3.0±0.30 2.7±0.10 2.6±0.30 ND ND + 0 3.9±0.03 3.4±0.11 2.9±0.29 ND 2.5±0.23 + 2 2.4±0.36 2.5±0.24 ND ND ND + 4 1.4±1.25 0.7±1.15 ND ND ND + 0 6.9±0.08 3.5±0.05 5.4±0.01 ND TNTC + 2 5.3±0.05 2.8±0.11 4.0±0.03 ND ND + Germinated 4 4.5±0.06 2.2±0.28 3.0±0.17 ND ND + brown rice 6 3.8±0.04 ND 1) 2.1±0.17 ND ND + 8 3.2±0.21 ND 0.8±1.33 ND ND + 10 2.3±0.24 ND 0.7±1.15 ND ND + Barley Soybean Oat 0 2.7±0.14 3.2±0.10 2.0±0.01 ND 1.3±0.24 + 2 ND ND ND ND ND + 0 4.5±0.03 3.0±0.19 3.4±0.03 ND TNTC + 2 2.8±0.32 2.6±0.28 2.4±0.10 ND ND + 4 2.1±0.17 1.4±1.25 ND ND ND + 6 ND ND ND ND ND + 0 2.8±0.16 2.4±0.43 ND 1) ND 0.8±0.68 + 2 2.0±0.01 1.4±1.25 ND ND ND + Values are mean±standard deviation (n=5). 1) Not detected within the detection limit 10 2 CFU g -1. 2) TNTC: too numberous to count. 3) Bacterial growth was detected after incubating 35 C for 48 h in a thioglycollate medium (positive) or not (negative).
시판분말원료의감마선위생화 33 Table 2. Effect of gamma irradiation on bacterial populations in powdered tubers and bacterial growth after incubation 35 C for 48 h Samples Potato Microbes (log CFU g -1 ) Irradiation dose (kgy) Total aerobic Bacillus Clostridium Coliform Bacterial Molds bacteria cereus perfringens bacteria growth 0 5.8±0.03 2.3±0.24 3.0±0.07 ND 2.5±0.04 + 2) 2 3.0±0.07 ND 0.7±1.15 ND ND + 4 1.3±1.15 ND ND ND ND + 6 ND 1) ND ND ND ND + 0 5.6±0.05 2.8±0.44 2.4±0.35 ND 1.9±0.33 + 2 3.4±0.07 2.6±0.19 0.7±1.15 ND ND + Jerusalem 4 3.0±0.07 2.6±0.24 0.7±1.15 ND ND + artichoke 6 2.3±0.30 1.5±1.31 0.7±1.15 ND ND + 8 0.7±1.15 ND ND ND ND + Sweet potato 0 6.0±0.16 2.7±0.09 5.4±0.01 ND 2.8±0.10 + 2 5.0±0.07 1.3±1.15 3.7±0.05 ND ND + 4 4.6±0.19 ND 2.5±0.15 ND ND + 6 4.4±0.03 ND 2.5±0.28 ND ND + 8 4.3±0.14 ND 2.1±0.17 ND ND + 10 4.2±0.06 ND 2.0±0.01 ND ND + Values are mean±standard deviation (n=5). 1) Not detected within the detection limit 10 2 CFU g -1. 2) Bacterial growth was detected after incubating 35 C for 48 h in a thioglycollate medium (positive) or not (negative). 3. 시판분말제품중엽채류분말의미생물평가시판분말제품중엽채류생식분말 3종 ( 샐러리, 케일및뽕잎 ) 의미생물오염수준과감마선조사처리된시료의일반호기성미생물, 진균류, B. cereus, 대장균군및 Cl. perfringens의생육정도및멸균여부확인결과는 Table 3에나타내었다. 조사처리되지않은샐러리, 케일및뽕잎분말에서일반호기성미생물은각각 5.9, 5.3 및 6.8 log CFU g -1 으로나타났다. Kim et al. (2012) 은케일의초기일반호기성미생물이 4.0 log CFU g -1 수준을나타냈으며 Koh and Kim (2010) 은 6.0 log CFU g -1 이었다고보고한바있다. 비조사구의진균류는샐러리, 케일및뽕잎분말에서각각 0.7, 2.9 및 2.8 log CFU g -1 으로확인되었으며, 대장균군의경우샐러리와케일은검출한계내에서 (2.0 log CFU g -1 ) 검출되지않았으나, 뽕잎분말에서는높은오염도를확인할수있었다. Cl. perfringens는엽채류분말 3종모두에서검출되지않았다 ( 검출한계 2.0 log CFU g -1 ). B. cereus의경우샐러리, 케일및뽕잎모두5.4 log CFU g -1 으로검출되었는데, 이는생식제품의미생물기준인 3.0 log CFU g -1 이상으로생식제품의기준에부적합한것으로확인되었다. 엽채류분말 3종모두4kGy 이하의선량으로감마선조사했을때식품공전의생식제품의기준에는충족 되었다. 그러나감마선조사된엽채류생식분말의멸균확인실험결과, 3종모두10 kgy에서멸균이되지않은것으로확인되었다. 엽채류와같은샐러리, 케일및뽕잎은엽육조직이얇고연약해조직파괴, 병균침입, 시듦등과같이외부환경에쉽게반응하게되므로신속하지않은제조공정은제품의초기오염도도높이고 (Jeong et al. 1996), 이로인해 10 kgy의조사선량으로도멸균되지않은것으로판단되었다. 결론적으로생식원료로사용가능한분말원료 14종중들깨, 메밀, 보리, 귀리, 감자, 돼지감자분말을제외한나머지 8종의분말제품들은식품공전의생식제품미생물기준을만족시키지못하여분말원료각각을생식으로제조할수는없었다. 다만시판되는생식제품들은다양한분말원료를혼합하기때문에이들의균수가희석되어그제품의미생물기준을충족시킬수는있을것으로판단된다. 그러나각각의분말원료의대장균군의수가높아음성이기준인대장균의검출위험이높으므로이에대한위생화방법이필수적으로고려되어야할것이다. 이에발아현미, 고구마및엽채류 3종을제외한나머지분말원료들은 10 kgy 조사선량에서멸균이가능했으며, 6 kgy의선량으로도식품공전의미생물기준을만족하는생식제품제조를위한위생화가가능하였다. 따라서본연구결과를통해가열살균이어려운분말
34 최수정 한인준 윤영민 김종헌 김재훈 김재경 박종흠 이주운 홍성길 육홍선 송범석 Table 3. Effect of gamma irradiation on bacterial populations in powdered leafy vegetables and bacterial growth after incubation 35 C for 48 h Samples Celery Kale Microbes (log CFU g -1 ) Irradiation dose (kgy) Total aerobic Bacillus Clostridium Coliform Bacterial Molds bacteria cereus perfringens bacteria growth 0 5.9±0.11 0.7±1.15 5.4±0.01 ND ND + 3) 2 4.1±0.03 ND 2.9±0.36 ND ND + 4 3.3±0.14 ND 2.6±0.51 ND ND + 6 3.1±0.08 ND 2.6±0.24 ND ND + 8 1.4±1.25 ND ND ND ND + 10 ND 1) ND ND ND ND + 0 5.3±0.02 2.9±0.03 5.4±0.01 ND ND + 2 4.3±0.09 2.5±0.21 3.7±0.06 ND ND + 4 3.5±0.07 1.6±1.38 2.9±0.14 ND ND + 6 2.7±0.34 0.7±1.15 2.3±0.30 ND ND + 8 2.5±0.45 ND ND ND ND + 10 2.4±0.10 ND ND ND ND + 0 6.8±0.11 2.8±0.14 5.4±0.01 ND TNTC 2) + 2 5.2±0.06 0.8±1.33 3.7±0.11 ND ND + Mulberry 4 4.6±0.16 ND 2.2±0.17 ND ND + leaves 6 4.3±0.04 ND 1.3±1.15 ND ND + 8 3.8±0.11 ND ND ND ND + 10 2.7±0.20 ND ND ND ND + Values are mean±standard deviation (n=5). 1) Not detected within the detection limit 10 2 CFU g -1. 2) TNTC: too numberous to count. 3) Bacterial growth was detected after incubating 35 C for 48 h in a thioglycollate medium (positive) or not (negative). Table 4. Microbiological specifications for Saeng-Sik (uncooked foods) products in the Korean Food Code Microbes Specification Bacillus cereus Not more than 1,000 CFU in 1 g of sample Clostridium perfringens Not more than 100 CFU in 1 g of sample Esherichia coli Negative 제품과가열을통한살균공정이없는생식제품의위생화를위한공정으로방사선기술이활용가능한것으로사료되었다. 보리, 귀리, 감자및돼지감자를제외한모든제품에서 Bacillus cereus가생식제품의기준인 3.0 log CFU g -1 을초과하였으나, 6 kgy의선량으로감마선조사처리시곡류, 서류및엽채류분말모두생식제품의미생물기준을만족하였다. 또한, 실험에사용된 14종의분말제품중발아현미, 고구마및엽채류 3종을제외한분말들은 10 kgy 이하의선량으로멸균이가능한것으로확인되었다. 사사 결론본연구는시판분말제품들을이용하여멸균생식제품을개발하기위해분말제품각각의미생물오염수준을확인한후위생화를위하여 1~10 kgy로감마선조사하여멸균효과를조사하였다. 실험에사용된분말은총14종으로곡류 8종, 서류 3종및엽채류 3종이었다. 비조사구에서곡류, 서류및엽채류분말의총균수는각각 2.7~6.9, 5.6~6.0 및 5.3~6.8 log CFU g -1 의수준으로나타났으며, 대장균군은율무, 기장, 발아현미, 백태및뽕잎분말에서높은오염도를보였다. 들깨, 메밀, 본연구는 2013년정부 ( 미래창조과학부 ) 의재원으로한국연구재단의지원및농림축산식품부고부가가치식품기술개발사업을통해수행되었으며, 그지원에감사드립니다. 참고문헌 김왕근, 정일윤, 이주운, 변명우. 2009. 방사선식품생명공학. 133-144pp. 1st ed. 도서출판생각나눔, 고양. Byun MW and Lee JW. 2003. Application of irradiation technology for food safety and security. Food Sci. Ind. 36:25-
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