Luận án Nghiên cứu tuyển chọn chủng nấm men để ứng dụng trong sản xuất whisky từ malt đại mạch và ngô

Luận án ―Nghiên cứu tuyển chọn chủng nấm men để ứng dụng trong sản xuất whisky từ malt đại mạch và ngô‖ đ giải quyết các vấn đề sau: 1. Từ nguồn bánh men truyền thống thu thập được ở ắc à (Lào ai), Mẫu Sơn (Lạng Sơn), làng Vân ( ắc iang) đ phân lập và tuyển chọn được 01 chủng nấm men (ký hiệu MS42). nghiên cứu đặc điểm hình thái, sinh lý, sinh h a và trình tự nucleotid của ADN, trên vùng ITS1, ITS2 của 5,8S và D1, D2 của 28S và xác định chủng MS42 thu c loài Saccharomyces cerevisiae và ký hiệu chủng là Saccharomyces cerevisiae MS42. 2. hủng MS42 c các đặc điểm sinh học sau: khả năng lên men dịch đường h a từ malt đại mạch và ngô tạo đ rượu đạt 12,3%Vol ở 25o , tạo được hương, vị tốt, đặc trưng cho sản ph m. hủng M42 c khả năng sinh trưởng tốt trên môi trường malt (M), ngô (N) sau 36 – 48 giờ nuôi cấy đạt số lượng tế bào tối đa từ 213 x 106 – 215 x 106 tế bào/ml. Lên men rượu tối ưu từ dịch lên men c hàm lượng đường:160 – 180g/l; pHopt: 5,0 – 5,5; Topt = 20 – 30o đạt đ cồn: 9,00 – 9,91%Vol. iệu suất lên men từ 89,52 – 91,13%, tạo ít đ c tố methanol, acetaldehyde, fufurol

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T i liệu t m k o tr n Internet 139. abed4070d3, ngày truy cập 10/9/2019. 140. https://www.businessinsider.com/the-biggest-whiskey-market-in-the-world- 2015-6, ngày truy cập 23/8/2019. 141. https://www.grandviewresearch.com/industry-analysis/travel-retail-market, ngày truy cập 28/8/2019. 142. https://tapchidouong.com.vn/nganh-bia-ruou-viet-nam-thuc-trang-san-xuat- kinh-doanh-va-su-dung-bia-ruou-1611.html, ngày truy cập 11/9/2019. 143. https://www.reportlinker.com/ci02016/Spirits.html, ngày truy cập 19/9/2019. 1PL 2PL PH C P ụ lục 1: T iết bị T thiết bị Nguồ gố PTN/ĐV - uồng đếm hồng cầu Thoma ức SP à N i - Máy đo đ cồn, chất hoà tan DA - 300 / KEM Density Nhật ản Cty Eresson - Máy đo p InoLab – WTW Nhật ản SP à N i - Máy ảnh Olympus 5050ZOOM Nhật ản SP à N i - Kính hiển vi điện tử Olympus X - 41 ức SP à N i - Kính hiển vi điện tử J M - T8 Nhật ản Viện VSDT TW - ân điện tử M – 200 Nhật SP à N i - Tủ sấy DKN - 811, Yamato Nhật ản SP à N i/ Cty Eresson - Nồi hấp SS325 - TOMY Mỹ SP à N i - Máy lắc SA - 32, Yamato Nhật SP à N i - Máy sắc ký khí System P 6890 series - t nhồi P - INNWax Polyethylene Glycol Italia Cty Eresson - hiết quang kế đo o x, đ cồn Trung Quốc SP à N i - chưng cất cồn SMI O Trung Quốc SP à N i - ệ thống chưng cất 20 lit; 100 lít Việt Nam SP à N i/ Cty Eresson - Máy nghiền uhley- Universal ức Cty Eresson - Máy đo nồng đ ôxy hoà tan ORION 5 Star Mỹ Cty Eresson - Tủ ổn nhiệt FO - 225E Italy SP à N i/ Cty Eresson 3PL P ụ lục 2: ôi trƣờn 2.1. M i trườ g giữ giố g a. Mô trườn Hansen [6] (ký ệu l : M1) Thành phần: Nước cất 1000,0 ml Cao men 1,0 g Pepton 10,0 g ường glucose 50,0 g MgSO4.7H2O 3,0 g Aga 20 g KH2PO4 3,0 g pH 6 - 7 b. Mô trườn dịc c ết malt - aga [6] (ký ệu l : M2) Thành phần: Dịch chiết malt (7 0Bx) 1000,0 ml NaCl 1,0 g KH2PO4 1,0 g Cao men 1,0 g MgSO4.7H2O 1,0 g Aga 20 g (NH4)2SO4 3,0 g pH 6 - 7 2.2. M i trườ g hâ si h hối ấm me YPD [5] Môi trường nhân sinh khối (ký hiệu là: M3) c thành phần như sau: Nước cất 1000,0 g Dextrose 20,0 g Cao men 10,0 g Aga 15 g Peptone 20,0 g pH 6,5 - 7,2 - C c mô trườn t an trùn ở 1210C /15 - 20 phút. 2.3. M i trườ g hâ giố g ơ b Môi trường nhân giống cơ bản (ký hiệu là: M6) c thành phần như sau: Dịch chiết malt 10 0Bx pH 5,0 g ách làm: oà tan các chất trong 1000 ml dịch chiết malt 100Bx. Thanh trùng ở 1150C trong 15 - 20 phút. 2.4. M i trườ g xá đị h h ă g si h trưở g, phát triể v me ủ á hủ g ấm me - M i trườ g hâ giố g ấp 1 ( ý hiệu : G1) 1- Pepton 10 (g/l) 3- Dịch malt 180Bx 20%Vol 2- Glucose 50 (g/l) 4- Nước (đủ tới) 1000 ml 3 – ường tổng số 80 (g/l) 5-p (chỉnh bằng citric) 4,5 - M i trườ g hâ giố g ơ b ( ý hiệu : G2) 1- Dịch malt 180Bx 50% V 3- ường tổng số 100 (g/l) 2- iá đ xay 100g/l 4-p (chỉnh bằng citric) 4,5 3 - Nước (đủ tới) 1000 ml 3PL P ụ lục 3: P ân lập nấm men ánh men ắc à Bánh men Làng Vân ánh men Mẫu Sơn ình 1: ình ảnh bánh men truyền thống của Việt Nam 4PL a. ánh giá hình thái khu n lạc b. MS18 d. MS42 e. MS18 g. LV22 c. BH30 ình 2: ình thái khu n lạc m t số mẫu nấm men điển hình 5PL a. Men ắc à b. Men Mẫu Sơn Men Làng Vân ình 3: Mẫu nấm men phân lập từ các nguồn khác nhau 6PL P ụ lục 4: N uy n liệu v tuyển c ọn nấm men a. Malt Buloke (Úc) b. Malt Sebastian (Pháp) c. Ngô DK 9955 ình 4: Mẫu malt, ngô nguyên liệu ình 5: Lên men các d ng nấm men phân lập trên môi trường malt, ngô 7PL a. Lên men của các d ng nấm men Mấu Sơn (MS) b. Lên men các d ng nấm men ắc à ( ) và LàngVân (LV) ình 6: ánh giá lên men của các d ng nấm men truyền thống 8PL P ụ lục 5: Đặc điểm l n men v đán iá c m qu n rƣợu từ các dòn nấm men ẫu Sơn ảng 1: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu malt các d ng nấm men Mẫu Sơn STT Mẫu mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 MS1 18 170,55 2,78 85,73 9,0 71,5 83,35 4,0 0,8 4,0 1,2 3,0 2,0 14,0 2 MS2 18 170,55 3,52 85,35 8,5 67,5 79,07 3,0 0,8 3,0 1,2 2,0 2,0 10,0 3 MS3 18 170,55 4,45 84,88 8,8 69,9 82,32 4,0 0,8 4,0 1,2 4,0 2,0 16,0 4 MS4 18 170,55 3,18 85,53 9,0 71,5 83,55 4,0 0,8 4,0 1,2 4,0 2,0 16,0 5 MS5 18 170,55 4,25 84,98 9,0 71,5 84,09 3,0 0,8 3,0 1,2 3,0 2,0 12,0 6 MS6 18 170,55 5,16 84,51 8,8 69,9 82,67 3,0 0,8 3,0 1,2 3,0 2,0 12,0 7 MS7 18 170,55 5,22 84,48 8,5 67,5 79,89 2,0 0,8 3,0 1,2 3,0 2,0 11,2 8 MS8 18 170,55 4,08 85,07 8,5 67,5 79,34 3,0 0,8 3,0 1,2 3,0 2,0 12,0 9 MS9 18 170,55 5,13 84,53 8,8 69,9 82,66 3,0 0,8 3,0 1,2 3,0 2,0 12,0 10 MS10 18 170,55 2,84 85,70 9,0 71,5 83,38 4,0 0,8 4,0 1,2 4,0 2,0 16,0 11 MS11 18 170,55 3,16 85,54 9,0 71,5 83,54 3,0 0,8 3,0 1,2 3,0 2,0 12,0 12 MS12 18 170,55 3,24 85,50 9,0 71,5 83,58 3,0 0,8 3,0 1,2 3,0 2,0 12,0 13 MS13 18 170,55 1,95 86,15 9,2 73,0 84,79 3,0 0,8 3,0 1,2 3,0 2,0 12,0 14 MS14 18 170,55 3,26 85,49 8,4 66,7 78,02 3,0 0,8 2,0 1,2 3,0 2,0 10,8 15 MS15 18 170,55 2,08 86,09 9,2 73,0 84,85 4,0 0,8 3,0 1,2 4,0 2,0 14,8 16 MS16 18 170,55 3,46 85,38 8,2 65,1 76,25 3,0 0,8 2,0 1,2 3,0 2,0 10,8 17 MS17 18 170,55 5,03 84,58 8,5 67,5 79,79 3,0 0,8 2,0 1,2 3,0 2,0 10,8 18 MS18 18 170,55 3,24 85,50 9,2 73,0 86,44 4,5 0,8 4,3 1,2 3,7 2,0 16,3 9PL 19 MS19 18 170,55 3,25 85,49 8,8 69,9 81,73 3,0 0,8 3,0 1,2 3,0 2,0 12,0 20 MS20 18 170,55 3,89 85,16 9,0 71,5 83,91 3,0 0,8 3,0 1,2 3,0 2,0 12,0 21 MS21 18 170,55 5,23 84,48 8,5 67,5 79,89 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 MS22 18 170,55 4,16 85,03 9,0 71,5 84,05 4,0 0,8 3,0 1,2 4,0 2,0 14,8 23 MS23 18 170,55 4,23 84,99 8,8 69,9 82,21 3,0 0,8 3,0 1,2 4,0 2,0 14,0 24 MS24 18 170,55 3,16 85,54 9,0 71,5 83,54 3,0 0,8 3,0 1,2 3,0 2,0 12,0 25 MS25 18 170,55 5,03 84,58 8,5 67,5 79,79 4,0 0,8 3,0 1,2 4,0 2,0 14,8 26 MS26 18 170,55 5,24 84,47 8,5 67,5 79,89 4,0 0,8 3,0 1,2 4,0 2,0 14,8 27 MS27 18 170,55 2,16 86,05 9,0 71,5 83,05 3,0 0,8 3,0 1,2 3,0 2,0 12,0 28 MS28 18 170,55 6,28 83,94 8,4 66,7 79,45 3,0 0,8 3,0 1,2 3,0 2,0 12,0 29 MS29 18 170,55 5,92 84,13 8,5 67,5 80,22 3,0 0,8 2,0 1,2 3,0 2,0 10,8 30 MS30 18 170,55 5,41 84,39 8,5 67,5 79,98 4,0 0,8 3,0 1,2 4,0 2,0 14,8 31 MS31 18 170,55 5,23 84,48 8,5 67,5 79,89 3,0 0,8 3,0 1,2 2,0 2,0 10,0 32 MS32 18 170,55 3,55 85,34 8,8 69,9 81,88 4,0 0,8 3,0 1,2 3,0 2,0 12,8 33 MS33 18 170,55 4,26 84,97 8,8 69,9 82,23 3,0 0,8 2,0 1,2 2,0 2,0 8,8 34 MS34 18 170,55 2,81 85,72 9,0 71,5 83,37 3,0 0,8 2,0 1,2 3,0 2,0 10,8 35 MS35 18 170,55 4,55 84,83 8,0 63,5 74,88 3,0 0,8 3,0 1,2 3,0 2,0 12,0 36 MS36 18 170,55 3,68 85,27 8,5 67,5 79,15 3,0 0,8 3,0 1,2 3,0 2,0 12,0 37 MS37 18 170,55 5,62 84,28 8,5 67,5 80,08 2,0 0,8 3,0 1,2 3,0 2,0 11,2 38 MS38 18 170,55 2,84 85,70 9,0 71,5 83,38 2,0 0,8 3,0 1,2 3,0 2,0 11,2 39 MS39 18 170,55 3,21 85,51 9,0 71,5 83,57 3,0 0,8 3,0 1,2 3,0 2,0 12,0 40 MS40 18 170,55 3,01 85,61 9,5 75,4 88,11 4,5 0,8 4,1 1,2 3,8 2,0 16,1 41 MS41 18 170,55 5,28 84,45 8,8 69,9 82,73 3,0 0,8 3,0 1,2 3,0 2,0 12,0 42 MS42 18 170,55 2,05 86,10 9,7 77,0 89,45 4,5 0,8 4,3 1,2 3,9 2,0 16,6 43 MS43 18 170,55 5,62 84,28 8,5 67,5 80,08 3,0 0,8 3,0 1,2 3,0 2,0 12,0 10PL ảng 2: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu ngô các d ng nấm men Mẫu Sơn STT Dòng mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 MS1 18 169,3 2,92 85,02 9,0 71,5 84,05 3,0 0,8 4,0 1,2 3,0 2,0 13,2 2 MS2 18 169,3 3,46 84,74 8,3 65,9 77,77 3,0 0,8 3,0 1,2 3,0 2,0 12,0 3 MS3 18 169,3 4,49 84,22 8,6 68,3 81,08 4,0 0,8 4,0 1,2 4,0 2,0 16,0 4 MS4 18 169,3 3,62 84,66 9,0 71,5 84,41 3,0 0,8 3,0 1,2 4,0 2,0 14,0 5 MS5 18 169,3 4,29 84,32 8,8 69,9 82,87 3,0 0,8 3,0 1,2 3,0 2,0 12,0 6 MS6 18 169,3 5,22 83,84 8,6 68,3 81,44 3,0 0,8 2,0 1,2 4,0 2,0 12,8 7 MS7 18 169,3 5,26 83,82 8,3 65,9 78,62 3,0 0,8 2,0 1,2 3,0 2,0 10,8 8 MS8 18 169,3 4,12 84,41 8,3 65,9 78,08 3,0 0,8 3,0 1,2 3,0 2,0 12,0 9 MS9 18 169,3 5,17 83,87 8,6 68,3 81,42 3,0 0,8 3,0 1,2 3,0 2,0 12,0 10 MS10 18 169,3 2,98 84,99 9,0 71,5 84,08 4,0 0,8 4,0 1,2 4,0 2,0 16,0 11 MS11 18 169,3 3,2 84,88 8,8 69,9 82,32 3,0 0,8 3,0 1,2 3,0 2,0 12,0 12 MS12 18 169,3 3,28 84,84 8,8 69,9 82,36 3,0 0,8 3,0 1,2 3,0 2,0 12,0 13 MS13 18 169,3 1,99 85,50 9,0 71,5 83,58 3,0 0,8 3,0 1,2 3,0 2,0 12,0 14 MS14 18 169,3 3,3 84,83 8,2 65,1 76,75 3,0 0,8 3,0 1,2 3,0 2,0 12,0 15 MS15 18 169,3 2,12 85,43 9,0 71,5 83,65 4,0 0,8 3,0 1,2 4,0 2,0 14,8 16 MS16 18 169,3 3,5 84,72 8,0 63,5 74,97 3,0 0,8 2,0 1,2 3,0 2,0 10,8 17 MS17 18 169,3 5,07 83,92 8,3 65,9 78,53 3,0 0,8 2,0 1,2 3,0 2,0 10,8 18 MS18 18 169,3 3,28 84,84 9,2 73,0 86,10 4,5 0,8 4,1 1,2 3,2 2,0 14,9 19 MS19 18 169,3 3,29 84,83 8,6 68,3 80,49 3,0 0,8 3,0 1,2 3,0 2,0 12,0 11PL 20 MS20 18 169,3 3,93 84,50 8,8 69,9 82,68 3,0 0,8 3,0 1,2 3,0 2,0 12,0 21 MS21 18 169,3 5,27 83,82 8,3 65,9 78,62 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 MS22 18 169,3 4,2 84,37 8,8 69,9 82,82 3,0 0,8 3,0 1,2 3,0 2,0 12,0 23 MS23 18 169,3 4,27 84,33 8,8 69,9 82,86 3,0 0,8 3,0 1,2 4,0 2,0 14,0 24 MS24 18 169,3 3,2 84,88 8,8 69,9 82,32 3,0 0,8 3,0 1,2 3,0 2,0 12,0 25 MS25 18 169,3 5,07 83,92 8,3 65,9 78,53 4,0 0,8 3,0 1,2 4,0 2,0 14,8 26 MS26 18 169,3 5,38 83,76 8,3 65,9 78,68 4,0 0,8 2,0 1,2 3,0 2,0 11,6 27 MS27 18 169,3 2,2 85,39 8,8 69,9 81,83 3,0 0,8 3,0 1,2 3,0 2,0 12,0 28 MS28 18 169,3 6,32 83,28 8,2 65,1 78,18 3,0 0,8 3,0 1,2 3,0 2,0 12,0 29 MS29 18 169,3 5,96 83,47 8,3 65,9 78,96 3,0 0,8 2,0 1,2 3,0 2,0 10,8 30 MS30 18 169,3 5,45 83,73 8,3 65,9 78,71 4,0 0,8 3,0 1,2 3,0 2,0 12,8 31 MS31 18 169,3 5,27 83,82 8,3 65,9 78,62 3,0 0,8 3,0 1,2 2,0 2,0 10,0 32 MS32 18 169,3 3,59 84,68 8,6 68,3 80,64 3,0 0,8 3,0 1,2 3,0 2,0 12,0 33 MS33 18 169,3 4,3 84,32 8,6 68,3 80,99 3,0 0,8 2,0 1,2 2,0 2,0 8,8 34 MS34 18 169,3 2,85 85,06 8,8 69,9 82,15 3,0 0,8 2,0 1,2 3,0 2,0 10,8 35 MS35 18 169,3 4,59 84,17 7,8 61,9 73,58 3,0 0,8 3,0 1,2 3,0 2,0 12,0 36 MS36 18 169,3 3,72 84,61 8,3 65,9 77,89 3,0 0,8 3,0 1,2 3,0 2,0 12,0 37 MS37 18 169,3 5,66 83,62 8,3 65,9 78,81 2,0 0,8 2,0 1,2 3,0 2,0 10,0 38 MS38 18 169,3 2,88 85,04 9,0 71,5 84,03 2,0 0,8 3,0 1,2 3,0 2,0 11,2 39 MS39 18 169,3 3,25 84,85 8,8 69,9 82,35 3,0 0,8 3,0 1,2 3,0 2,0 12,0 40 MS40 18 169,3 3,05 84,95 9,4 74,6 87,85 4,5 0,8 4,1 1,2 3,2 2,0 14,9 41 MS41 18 169,3 5,32 83,79 8,6 68,3 81,49 3,0 0,8 3,0 1,2 3,0 2,0 12,0 42 MS42 18 169,3 2,16 85,41 9,5 75,4 88,32 4,8 0,8 4,2 1,2 3,5 2,0 15,9 43 MS43 18 169,3 5,66 83,62 8,5 67,5 80,71 3,0 0,8 2,0 1,2 3,0 2,0 10,8 12PL P ụ lục 6: Đặc điểm l n men v đán iá c m qu n rƣợu từ các dòn nấm men ắc H ảng 3: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu malt các d ng nấm men ắc à STT Dòng mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 BH1 18 170,55 3,24 85,50 9,0 71,5 83,58 4,3 0,8 3,1 1,2 3,9 2,0 15,0 2 BH2 18 170,55 2,95 85,64 9,0 71,5 83,44 4,6 0,8 3,9 1,2 3,7 2,0 15,8 3 BH3 18 170,55 3,26 85,49 9,0 71,5 83,59 4,0 0,8 4,0 1,2 3,0 2,0 14,0 4 BH4 18 170,55 2,08 86,09 9,0 71,5 83,01 4,0 0,8 4,0 1,2 3,0 2,0 14,0 5 BH5 18 170,55 3,46 85,38 9,0 71,5 83,69 3,0 0,8 3,0 1,2 3,0 2,0 12,0 6 BH6 18 170,55 5,03 84,58 8,8 69,9 82,61 3,0 0,8 3,0 1,2 3,0 2,0 12,0 7 BH7 18 170,55 3,24 85,50 9,0 71,5 83,58 2,0 0,8 3,0 1,2 3,0 2,0 11,2 8 BH8 18 170,55 3,25 85,49 9,0 71,5 83,59 3,0 0,8 3,0 1,2 3,0 2,0 12,0 9 BH9 18 170,55 3,89 85,16 8,8 69,9 82,04 3,0 0,8 4,0 1,2 3,0 2,0 13,2 10 BH10 18 170,55 5,23 84,48 8,8 69,9 82,71 3,0 0,8 4,0 1,2 4,0 2,0 15,2 11 BH11 18 170,55 4,16 85,03 8,5 67,5 79,38 3,0 0,8 3,0 1,2 3,0 2,0 12,0 12 BH12 18 170,55 3,24 85,50 8,8 69,9 81,73 3,0 0,8 3,0 1,2 3,0 2,0 12,0 13 BH13 18 170,55 1,95 86,15 9,2 73,0 84,79 3,0 0,8 3,0 1,2 3,0 2,0 12,0 14 BH14 18 170,55 3,26 85,49 8,4 66,7 78,02 3,0 0,8 2,0 1,2 3,0 2,0 10,8 15 BH15 18 170,55 2,08 86,09 9,2 73,0 84,85 4,0 0,8 3,0 1,2 4,0 2,0 14,8 16 BH16 18 170,55 3,46 85,38 8,2 65,1 76,25 3,0 0,8 2,0 1,2 3,0 2,0 10,8 17 BH17 18 170,55 5,03 84,58 8,5 67,5 79,79 3,0 0,8 2,0 1,2 3,0 2,0 10,8 18 BH18 18 170,55 3,24 85,50 9,0 71,5 83,58 3,0 0,8 4,0 1,2 4,0 2,0 15,2 19 BH19 18 170,55 3,25 85,49 9,5 75,4 88,23 4,5 0,8 3,9 1,2 3,9 2,0 16,1 13PL 20 BH20 18 170,55 2,16 86,05 8,8 69,9 81,20 4,1 0,8 3,1 1,2 4,2 2,0 15,4 21 BH21 18 170,55 6,28 83,94 8,5 67,5 80,40 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 BH22 18 170,55 5,92 84,13 8,5 67,5 80,22 4,0 0,8 3,0 1,2 4,0 2,0 14,8 23 BH23 18 170,55 5,41 84,39 9,0 71,5 84,68 3,0 0,8 3,0 1,2 4,0 2,0 14,0 24 BH24 18 170,55 5,23 84,48 9,2 73,0 86,47 3,5 0,8 4,1 1,2 3,9 2,0 15,5 25 BH25 18 170,55 3,55 85,34 8,5 67,5 79,09 4,0 0,8 3,0 1,2 4,0 2,0 14,8 26 BH26 18 170,55 4,26 84,97 9,0 71,5 84,10 4,0 0,8 3,0 1,2 4,0 2,0 14,8 27 BH27 18 170,55 2,81 85,72 9,0 71,5 83,37 3,0 0,8 3,0 1,2 3,0 2,0 12,0 28 BH28 18 170,55 4,55 84,83 8,4 66,7 78,63 3,0 0,8 3,0 1,2 3,0 2,0 12,0 29 BH29 18 170,55 3,68 85,27 8,5 67,5 79,15 3,0 0,8 2,0 1,2 3,0 2,0 10,8 30 BH30 18 170,55 5,62 84,28 9,0 71,5 84,79 4,6 0,8 4,1 1,2 3,8 2,0 16,2 31 BH31 18 170,55 2,84 85,70 8,5 67,5 78,75 3,0 0,8 3,0 1,2 2,0 2,0 10,0 32 BH32 18 170,55 3,21 85,51 8,8 69,9 81,71 4,0 0,8 3,0 1,2 3,0 2,0 12,8 33 BH33 18 170,55 3,01 85,61 9,5 75,4 88,11 4,2 0,8 3,4 1,2 4,2 2,0 15,8 34 BH34 18 170,55 5,28 84,45 9,0 71,5 84,62 4,1 0,8 3,2 1,2 4,1 2,0 15,3 35 BH35 18 170,55 4,55 84,83 9,0 71,5 84,24 3,0 0,8 3,0 1,2 3,0 2,0 12,0 36 BH36 18 170,55 3,68 85,27 9,0 71,5 83,80 3,0 0,8 3,0 1,2 3,0 2,0 12,0 37 BH37 18 170,55 5,62 84,28 8,5 67,5 80,08 2,0 0,8 3,0 1,2 3,0 2,0 11,2 38 BH38 18 170,55 2,84 85,70 8,5 67,5 78,75 2,0 0,8 3,0 1,2 3,0 2,0 11,2 39 BH39 18 170,55 3,21 85,51 8,8 69,9 81,71 3,0 0,8 3,0 1,2 3,0 2,0 12,0 40 BH40 18 170,55 3,01 85,61 8,8 69,9 81,61 3,0 0,8 4,0 1,2 4,0 2,0 15,2 41 BH41 18 170,55 5,28 84,45 8,5 67,5 79,91 3,9 0,8 3,1 1,2 4,3 2,0 15,4 42 BH42 18 170,55 2,95 85,64 9,2 73,0 85,29 4,2 0,8 3,8 1,2 3,6 2,0 15,2 14PL ảng 4: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu ngô các d ng nấm men ắc à STT Dòng mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 BH1 18 169,3 3,64 84,65 9,0 71,5 84,42 4,2 0,8 4,2 1,2 3,2 2,0 14,8 2 BH2 18 169,3 2,15 85,41 9,0 71,5 83,66 4,2 0,8 4,1 1,2 3,1 2,0 14,5 3 BH3 18 169,3 3,66 84,64 8,0 63,5 75,05 4,0 0,8 4,0 1,2 3,0 2,0 14,0 4 BH4 18 169,3 2,48 85,25 7,8 61,9 72,65 4,0 0,8 3,0 1,2 3,0 2,0 12,8 5 BH5 18 169,3 3,86 84,54 7,8 61,9 73,26 4,0 0,8 3,0 1,2 3,0 2,0 12,8 6 BH6 18 169,3 5,43 83,74 7,6 60,3 72,06 4,0 0,8 3,0 1,2 3,0 2,0 12,8 7 BH7 18 169,3 3,84 84,55 8,0 63,5 75,13 4,0 0,8 3,0 1,2 3,0 2,0 12,8 8 BH8 18 169,3 3,55 84,70 7,8 61,9 73,12 4,0 0,8 3,0 1,2 3,0 2,0 12,8 9 BH9 18 169,3 4,29 84,32 7,6 60,3 71,57 4,0 0,8 3,0 1,2 3,0 2,0 12,8 10 BH10 18 169,3 5,63 83,64 7,6 60,3 72,15 4,0 0,8 4,0 1,2 3,0 2,0 14,0 11 BH11 18 169,3 4,56 84,18 7,3 58,0 68,85 4,0 0,8 3,0 1,2 3,0 2,0 12,8 12 BH12 18 169,3 3,64 84,65 7,6 60,3 71,28 4,0 0,8 3,0 1,2 3,0 2,0 12,8 13 BH13 18 169,3 2,35 85,31 8,0 63,5 74,46 4,0 0,8 3,0 1,2 3,0 2,0 12,8 14 BH14 18 169,3 3,66 84,64 7,2 57,2 67,54 4,0 0,8 2,0 1,2 3,0 2,0 11,6 15 BH15 18 169,3 2,48 85,25 9,2 73,0 85,69 4,0 0,8 3,0 1,2 4,0 2,0 14,8 16 BH16 18 169,3 3,86 84,54 7,0 55,6 65,74 4,0 0,8 2,0 1,2 3,0 2,0 11,6 17 BH17 18 169,3 5,43 83,74 7,3 58,0 69,22 4,0 0,8 2,0 1,2 3,0 2,0 11,6 18 BH18 18 169,3 3,64 84,65 7,8 61,9 73,16 4,0 0,8 4,0 1,2 4,0 2,0 16,0 19 BH19 18 169,3 3,05 84,65 9,00 71,5 87,42 4,1 0,8 4,0 1,2 3,5 2,0 15,1 20 BH20 18 169,3 2,56 85,20 8,50 67,5 79,21 3,6 0,8 4,3 1,2 3,1 2,0 14,2 15PL 21 BH21 18 169,3 6,68 83,10 7,3 58,0 69,75 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 BH22 18 169,3 6,52 83,18 7,3 58,0 69,68 4,0 0,8 3,0 1,2 4,0 2,0 14,8 23 BH23 18 169,3 5,81 83,54 7,8 61,9 74,13 3,0 0,8 3,0 1,2 4,0 2,0 14,0 24 BH24 18 169,3 5,63 83,64 8,50 67,5 80,70 3,2 0,8 4,1 1,2 3,6 2,0 14,7 25 BH25 18 169,3 3,95 84,49 7,5 59,6 70,48 4,0 0,8 3,0 1,2 4,0 2,0 14,8 26 BH26 18 169,3 4,56 84,18 7,8 61,9 73,57 4,0 0,8 3,0 1,2 4,0 2,0 14,8 27 BH27 18 169,3 3,21 84,87 7,8 61,9 72,97 3,0 0,8 3,0 1,2 3,0 2,0 12,0 28 BH28 18 169,3 4,95 83,98 7,2 57,2 68,07 3,0 0,8 3,0 1,2 3,0 2,0 12,0 29 BH29 18 169,3 4,08 84,43 7,3 58,0 68,65 3,0 0,8 2,0 1,2 3,0 2,0 10,8 30 BH30 18 169,3 4,02 84,46 9,00 71,5 84,61 4,5 0,8 4,1 1,2 3,5 2,0 15,5 31 BH31 18 169,3 3,24 84,86 7,3 58,0 68,31 3,0 0,8 3,0 1,2 3,0 2,0 12,0 32 BH32 18 169,3 3,61 84,67 7,6 60,3 71,27 4,0 0,8 3,0 1,2 3,0 2,0 12,8 33 BH33 18 169,3 3,41 84,77 9,30 73,8 87,11 3,9 0,8 4,2 1,2 3,2 2,0 14,6 34 BH34 18 169,3 3,28 84,84 9,00 71,5 84,23 4,2 0,8 4,1 1,2 3,2 2,0 14,7 35 BH35 18 169,3 4,95 83,98 7,8 61,9 73,74 3,0 0,8 3,0 1,2 3,0 2,0 12,0 36 BH36 18 169,3 4,08 84,43 7,8 61,9 73,36 3,0 0,8 3,0 1,2 3,0 2,0 12,0 37 BH37 18 169,3 6,02 83,44 7,3 58,0 69,47 2,0 0,8 3,0 1,2 3,0 2,0 11,2 38 BH38 18 169,3 3,24 84,86 7,3 58,0 68,31 2,0 0,8 3,0 1,2 3,0 2,0 11,2 39 BH39 18 169,3 3,85 84,54 7,6 60,3 71,38 3,0 0,8 3,0 1,2 3,0 2,0 12,0 40 BH40 18 169,3 3,41 84,77 7,6 60,3 71,19 4,0 0,8 3,0 1,2 4,0 2,0 14,8 41 BH41 18 169,3 5,68 83,61 8,50 67,5 80,72 3,9 0,8 4,1 1,2 3,5 2,0 15,0 42 BH42 18 169,3 3,05 84,95 9,00 71,5 84,12 4,3 0,8 3,8 1,2 3,2 2,0 14,4 16PL P ụ lục 7: Đặc điểm l n men v đán iá c m qu n rƣợu từ các dòn nấm men Làng Vân ảng 5: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu malt các d ng nấm men Làng Vân STT Dòng mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 LV1 18 170,55 4,24 84,98 7,8 61,9 72,87 3,5 0,8 4,0 1,2 3,0 2,0 13,6 2 LV2 18 170,55 2,68 85,78 8,0 63,5 74,05 3,5 0,8 3,5 1,2 3,0 2,0 13,0 3 LV3 18 170,55 3,26 85,49 8,0 63,5 74,31 3,2 0,8 3,0 1,2 4,0 2,0 14,2 4 LV4 18 170,55 2,08 86,09 8,5 67,5 78,40 3,6 0,8 3,9 1,2 3,7 2,0 15,0 5 LV5 18 170,55 3,46 85,38 7,8 61,9 72,53 3,0 0,8 3,0 1,2 3,0 2,0 12,0 6 LV6 18 170,55 5,03 84,58 7,5 59,6 70,41 3,5 0,8 3,0 1,2 3,0 2,0 12,4 7 LV7 18 170,55 4,24 84,98 8,0 63,5 74,74 3,5 0,8 3,2 1,2 3,0 2,0 12,6 8 LV8 18 170,55 3,25 85,49 8,0 63,5 74,30 3,5 0,8 3,0 1,2 3,0 2,0 12,4 9 LV9 18 170,55 3,89 85,16 7,8 61,9 72,72 3,0 0,8 3,0 1,2 3,0 2,0 12,0 10 LV10 18 170,55 5,23 84,48 7,5 59,6 70,49 3,5 0,8 3,0 1,2 4,0 2,0 14,4 11 LV11 18 170,55 4,16 85,03 7,8 61,9 72,84 3,5 0,8 3,0 1,2 3,0 2,0 12,4 12 LV12 18 170,55 3,24 85,50 8,2 65,1 76,15 3,0 0,8 3,0 1,2 3,0 2,0 12,0 13 LV13 18 170,55 1,95 86,15 8,2 65,1 75,57 3,2 0,8 3,0 1,2 3,0 2,0 12,2 14 LV14 18 170,55 3,26 85,49 7,4 58,8 68,73 3,2 0,8 3,0 1,2 3,0 2,0 12,2 15 LV15 18 170,55 2,08 86,09 8,2 65,1 75,63 3,0 0,8 3,0 1,2 4,0 2,0 14,0 17PL 16 LV16 18 170,55 4,46 84,87 7,2 57,2 67,36 3,5 0,8 3,0 1,2 3,0 2,0 12,4 17 LV17 18 170,55 5,03 84,58 7,5 59,6 70,41 3,5 0,8 3,0 1,2 3,0 2,0 12,4 18 LV18 18 170,55 3,25 85,49 8,2 65,1 76,16 3,5 0,8 3,0 1,2 4,0 2,0 14,4 19 LV19 18 170,55 3,25 85,49 8,5 67,5 78,94 3,5 0,8 4,0 1,2 3,5 2,0 14,6 20 LV20 18 170,55 2,15 86,05 7,8 61,9 71,97 3,2 0,8 3,0 1,2 3,0 2,0 12,2 21 LV21 18 170,55 6,28 83,94 7,5 59,6 70,94 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 LV22 18 170,55 3,26 85,49 9,2 73,0 85,45 4,6 0,8 4,2 1,2 3,8 2,0 16,3 18PL ảng 6: Tổng hợp lên men và đánh giá cảm quan rượu từ nguyên liệu ngô các d ng nấm men Làng Vân STT Dòng mấm men o Bx ường khử (quy về maltose) g/l ường sót (quy về maltose) g/l Khối lượng rượu theo lý thuyết (g) Phần trăm cồn thực tế (% v/v) Khối lượng rượu theo thực tế (g) iệu suất lên men rượu (%) ánh giá cảm quan (T VN 3215-79) trong và màu sắc ệ số (0,8) Mùi ệ số (1,2) Vị ệ số (2,0) Tổng điểm 1 LV1 18 169,3 4,54 84,19 7,8 61,9 73,56 4,0 0,8 4,0 1,2 3,0 2,0 14,0 2 LV2 18 169,3 3,28 84,84 7,0 55,6 65,51 3,0 0,8 4,0 1,2 3,0 2,0 13,2 3 LV3 18 169,3 3,46 84,74 7,8 61,9 73,08 4,0 0,8 4,0 1,2 3,0 2,0 14,0 4 LV4 18 169,3 2,58 85,19 8,5 67,5 79,22 4,5 0,8 4,1 1,2 3,2 2,0 14,9 5 LV5 18 169,3 3,96 84,49 7,6 60,3 71,42 3,0 0,8 3,0 1,2 3,0 2,0 12,0 6 LV6 18 169,3 5,23 83,84 7,3 58,0 69,13 3,0 0,8 3,0 1,2 3,0 2,0 12,0 7 LV7 18 169,3 4,74 84,09 8,0 63,5 75,54 2,0 0,8 3,0 1,2 3,0 2,0 11,2 8 LV8 18 169,3 3,75 84,60 7,8 61,9 73,21 3,0 0,8 3,0 1,2 3,0 2,0 12,0 9 LV9 18 169,3 4,59 84,17 7,6 60,3 71,70 3,0 0,8 3,0 1,2 3,0 2,0 12,0 10 LV10 18 169,3 5,73 83,58 7,3 58,0 69,35 4,0 0,8 4,0 1,2 3,0 2,0 14,0 11 LV11 18 169,3 4,66 84,13 7,6 60,3 71,73 3,0 0,8 3,0 1,2 3,0 2,0 12,0 12 LV12 18 169,3 3,54 84,70 8,0 63,5 74,99 3,0 0,8 3,0 1,2 3,0 2,0 12,0 13 LV13 18 169,3 2,45 85,26 8,0 63,5 74,50 3,0 0,8 3,0 1,2 3,0 2,0 12,0 14 LV14 18 169,3 3,76 84,59 7,2 57,2 67,58 3,0 0,8 2,0 1,2 3,0 2,0 10,8 15 LV15 18 169,3 2,58 85,19 8,0 63,5 74,56 4,0 0,8 3,0 1,2 4,0 2,0 14,8 16 LV16 18 169,3 4,96 83,98 8,0 63,5 75,64 3,0 0,8 3,0 1,2 3,0 2,0 12,0 19PL 17 LV17 18 169,3 5,53 83,69 7,5 59,6 71,16 3,0 0,8 3,0 1,2 3,0 2,0 12,0 18 LV18 18 169,3 3,55 84,70 7,8 61,9 73,12 4,0 0,8 4,0 1,2 3,0 2,0 14,0 19 LV19 18 169,3 3,75 84,60 8,3 65,9 77,90 5,0 0,8 5,0 1,2 4,0 2,0 18,0 20 LV20 18 169,3 2,65 85,16 8,5 67,5 79,25 3,0 0,8 3,0 1,2 3,0 2,0 12,0 21 LV21 18 169,3 6,78 83,05 8,3 65,9 79,35 4,0 0,8 3,0 1,2 4,0 2,0 14,8 22 LV22 18 169,3 3,26 84,85 9,0 71,5 84,22 4,5 0,8 3,9 1,2 3,5 2,0 15,3 20PL P ụ lục 8: Hìn n tế b o nấm men s u tuyển c ọn l n 1 BH1 BH2 BH19 BH20 BH24 BH30 BH33 BH34 BH41 BH42 LV4 LV22 MS18 MS40 MS42 ình 5: ình ảnh tế bào 15 chủng nấm men đ tuyển chọn lần 1 21PL P ụ lục 9: ết qu kiểm tr kit t ử k n n sử dụn đƣờn củ S42 a. Kết quả xác định khả năng sử dụng đường của chủng MS42 với kit ID 32 b. Kết quả xác định khả năng sử dụng đường của chủng MS42 với kit API 20 AUX ình 7. Kết quả kit thử khả năng sử dụng đường của chủng MS42 22PL P ụ lục 10: Các mẫu ỗ sồi sử dụn tron n i n cứu s n xuất rƣợu w isky từ n ô v m lt đ i m c ình 7: ác mẫu g sồi Mỹ 23PL ình 8: ác mẫu g sồi Pháp ình 9: ác mẫu g sồi Australia 24PL P ụ lục 11: ột số ìn n s n xuất rƣợu t i n máy bi rƣợu resson Máy nghiền Nồi đường h a ệ thống làm lạnh Tank nhân giống Tank lên men Thiết bị chưng cất chân không ệ thống điều khiển Kiểm tra chất lượng nấm men Lấy mẫu rượu phân tích ình 6: M t số hình ảnh sản xuất thực nghiệm tại nhà máy rượu bia RISSON P ụ lục 12: Sắc ký đồ các mẫu rƣợu trắn trƣớc k i n âm ỗ sồi 1. Mẫu M 100 (6/05/2016) 2. Mẫu N 00 (6/05/2016) 3. Mẫu 8N:2M (6/05/2016) P ụ lục 13: Sắc ký đồ các mẫu rƣợu n âm ỗ sồi 06 t án (S8.M, S8.N, S8.N8:M2) 1. Mẫu S8.M (5/11/2016) 2. Mẫu S8.N8:M2 (5/11/2016) 3. Mẫu: S8.N (5/11/2016) P ụ lục 14: Sắc ký đồ các mẫu rƣợu n âm ỗ sồi 12 t án 1. Mẫu S8.M (10/05/2017) 2. Mẫu S8.N (6/05/2017) 3. Mẫu S8. N8:M2 (10/05/2017) P ụ lục 15: Sắc ký đồ các mẫu rƣợu n âm ỗ sồi 18 t án 1. Mẫu: S8.M (10/11/2017) 2. Mẫu: S8.N (6/11/2017) 3. Mẫu S8. N8:M2 (10/11/2017) P ụ lục 16: Sắc ký đồ các mẫu rƣợu n âm ỗ sồi 24 t án 1. Mẫu: S8.M (2/05/2018) 2. Mẫu S8.N (2/05/2018) 4. Mẫu: S8.N8:M2 (2/05/2018)

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