Malolactic bacteria 고정화 오크칩을 활용한 Muscat Bailey A 레드와인의 malolactic fermentation 최적 조건 및 품질 특성 = Optimal Conditions and Quality Characteristics of Malolactic Fermentation of Muscat Bailey A Red Wine Using Malolactic Bacteria Immobilized Oak Chips

다국어 초록 (Multilingual Abstract)

This study was conducted to improve the sensory properties such as taste and flavor in wine brewing using domestic grape varieties, and investigated to the optimal condition of malolactic fermentation (MLF). In addition, the possibility of simultaneous progression of MLF and oak aging of Muscat Bailey A (MBA) wine was investigated using a technique to immobilize malolactic bacteria (MLB) causing MLF on oak chips.
The MLB for the progression of MLF was selected from L. plantarum JH287 with the highest acid resistance and O. oeni KCTC3091 with the highest alcohol resistance in environmental resistance tests such as sulfurous acid resistance, alcohol resistance, heat resistance, sugar resistance and acid resistance. two selected strains were inoculated into grape juice prior to alcoholic fermentation (AF) at a constant concentration (1×106, 1×107, 1×108 CFU/mL) to perform MLF. also, at the time when the malic acid content was the lowest the quality characteristics of wine prepared by sequential AF by inoculation with S. cerevisiae Fermivin were investigated. as a result, when inoculated with L. plantarum JH287 at 1×108 CFU/mL concentration, all of 1.06±0.02 mg/mL malic acid was converted to 0.76±0.02 mg/mL lactic acid within 24 hours after inoculation and this was the shortest with about 11 days. However, inoculation with O. oeni KCTC3091 showed little MLF activity at all inoculation concentrations, 0.97±0.02 mg/mL malic acid remained and only 0.27±0.02 mg/mL lactic acid was produced. through this, the concentration of 1×108 CFU/mL showing the shortest MLF time was established as the optimal MLF inoculation concentration, and the quality characteristics of wine prepared by inoculating two strains after AF according to the established optimal inoculation concentration were examined. as a result, when inoculated with L. plantarum JH287 at 1×108 CFU/mL 1.20±0.02 mg/mL of malic acid was converted to 0.18±0.04 mg/mL of lactic acid and 0.79±0.02 mg/mL of malic acid was remained. On the other hand, when inoculated with O. oeni KCTC3091 0.31±0.03 mg/mL of lactic acid was produced and 0.57±0.02 mg/mL of malic acid remained which was 1.5 times higher than that of the former.
American oak chips (Quercus alba) and french oak chips (Quercus robur) were explosion treatment at a constant temperature (160℃, 180℃, 200℃) and time (3 min, 4 min, 5 min). as a result, the conditions of 180℃ and 4 min, which showed the best appearance for the field emission scanning electron microscope (FE-SEM) observation, were established as the optimum processing conditions for oak chip explosion. when the degree of pore formed on the surface of two types of oak chips were examined using FE-SEM, american oak chips having relatively constant pore were finally selected as an MLB immobilized support.
L. plantarum JH287 and O. oeni KCTC3091 were immobilized on exploded oak chips and unexploded oak chips, respectively. in case of L. plantarum JH287 showed 8.9 log CFU/mL for the exploded oak chips, and 7.7 log CFU/mL for the unexploded oak chips. also in case of O. oeni KCTC3091 showed 8.9 log CFU/mL for the exploded oak chips, and 6.6 log CFU/mL for the unexploded oak chips. In addition, MLB immobilized oak chips were stored at 4℃. and 20℃. for 120 days, and the survival rate of the immobilized MLB was calculated for each period. As a result, L. plantarum JH287 and O. oeni KCTC3091 showed a survival rate of over 90% for up to 60 days when stored at 4℃. On the other hand, when stored at 20℃, a sudden decrease in survival rate appeared from 15 days.
Finally, MBA wine was prepared using MLB immobilized oak chips. As a result, sugar and reducing sugar decreased as fermentation progressed, and after completion of fermentation, sugar content ranged from 6.1~6.2 °Brix and reducing sugar was 0.14~0.25%, similar concentration under all conditions showed. In addition, the total acid finally showed a content of 0.72±0.03%, and showed similar concentration under all conditions. when fermented by O. oeni KCTC3091 culture medium (Sc-Oo-Oak) and immobilized oak chip (Sc-(i)Oo), pH was finally 3.62±0.04. also, fermented by L. plantarum JH287 culture medium (Lp-Sc-Oak) and immobilized oak chips ((i)Lp-Sc), pH was finally 3.50±0.01. The final alcohol content was 11.6±0.4% at all conditions and showed the general alcohol content. In case of Lp-Sc-Oak and (i)Lp-Sc was increased of the organic acid content of acetic acid. so, In sensory evaluation, the color score of (i)Lp-Sc received the highest score, but (i)Lp-Sc and Lp-Sc-Oak scored highest in sour taste and the lowest score in overall preference. On the other hand, Sc-Oo-Oak and Sc-(i)Oo scored the lowest in sour taste and Sc-(i)Oo scored the highest in overall preference.

목차 (Table of Contents)

• Ⅰ. 서론 1
• Ⅱ. 재료 및 방법 4
• 1. 실험재료 및 균주 배양 조건 4
• 가. 사용 균주 및 배양 조건 4
• 나. 균주의 환경내성 5
• 1) 아황산 내성 분석 5
• 2) 알코올 내성 분석 5
• 3) 내열성 분석 5
• 4) 내당성 분석 5
• 5) 내산성 분석 6
• 다. 실험재료 6
• 2. Malolactic bacteria 접종 시기에 따른 품질 특성 분석 7
• 가. 당도 및 알코올 함량 분석 7
• 나. pH 및 총산 분석 7
• 다. 생균수 분석 8
• 라. Malic acid 및 lactic acid 함량 분석 8
• 3. 오크칩 폭쇄 9
• 가. 폭쇄 처리 조건 및 오크칩 선정 9
• 나. 폭쇄 오크칩 제조 9
• 4. Malolactic bacteria 고정화 및 동결건조 10
• 가. 균체 획득 10
• 나. Protectant 조성 및 처리 10
• 다. Malolactic bacteria의 오크칩 고정화 10
• 라. 동결건조 처리 및 저장 11
• 5. 전계방사형 주사전자현미경(Field Emission Scanning Electron Microscope, FE-SEM) 관찰 및 생존율 측정 12
• 가. 고정화 오크칩의 FE-SEM 관찰 12
• 나. 고정화 오크칩의 malolactic bacteria 생존율 측정 12
• 6. 포도주 제조 13
• 7. 포도주의 발효 및 이화학적 특성 분석 15
• 가. 당도 및 환원당 함량 분석 15
• 나. pH 및 총산 분석 15
• 다. 알코올 함량 분석 15
• 라. 생균수 분석 16
• 마. 항산화 성분 함량 분석 16
• 1) 총 페놀성 화합물 함량 분석 16
• 2) 총 플라보노이드 함량 분석 16
• 3) 총 안토시아닌 함량 분석 16
• 바. 항산화 활성 분석 17
• 1) DPPH 라디칼 소거 활성 분석 17
• 2) ABTS 라디칼 소거 활성 분석 17
• 3) FRAP (Ferric ion Reducing Antioxidant Power) 분석 18
• 사. Malic acid 및 lactic acid 함량 분석 18
• 아. 유기산 및 유리당 조성 분석 18
• 자. Hue & Intensity 및 색도 분석 19
• 8. 관능평가 20
• 9. 통계처리 20
• Ⅲ. 결과 및 고찰 21
• 1. 균주의 선정 21
• 가. 균주의 환경내성 21
• 1) 아황산 내성 21
• 2) 알코올 내성 23
• 3) 내열성 25
• 4) 내당성 26
• 5) 내산성 28
• 2. Malolactic bacteria 접종 시기에 따른 품질 특성 30
• 가. Alcoholic fermentation 이전의 malolactic fermentation 30
• 1) 당도 및 알코올 함량 변화 30
• 2) pH 및 총산 변화 32
• 3) 생균수 변화 34
• 4) Malic acid 및 lactic acid 함량 변화 35
• 나. Alcoholic fermentation 이후의 malolactic fermentation 37
• 1) 당도 및 알코올 함량 변화 37
• 2) pH 및 총산 변화 38
• 3) 생균수 변화 39
• 4) Malic acid 및 lactic acid 함량 변화 40
• 3. 오크칩 폭쇄 및 선정 41
• 가. 처리 조건에 따른 오크칩의 외형 변화 41
• 나. 오크칩 최종 선정 43
• 4. 고정화 오크칩의 FE-SEM 관찰 44
• 5. 고정화 오크칩의 malolactic bacteria 생존율 변화 46
• 6. 포도주의 발효 및 이화학적 특성 48
• 가. 당도 및 환원당 함량 변화 48
• 나. pH 및 총산 변화 49
• 다. 알코올 함량 변화 50
• 라. 생균수 변화 50
• 마. 항산화 성분 함량 52
• 1) 총 페놀성 화합물 함량 52
• 2) 총 플라보노이드 함량 53
• 3) 총 안토시아닌 함량 54
• 바. 항산화 활성 55
• 1) DPPH 라디칼 소거 활성 55
• 2) ABTS 라디칼 소거 활성 56
• 3) FRAP (Ferric ion Reducing Antioxidant Power) 57
• 사. Malic acid 및 lactic acid 함량 변화 58
• 아. 유기산 및 유리당 조성 59
• 자. Hue & Intensity 및 색도 61
• 7. 관능평가 62
• Ⅳ. 요약 64
• Ⅴ. 참고문헌 67
• 영문초록 74