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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ang Thảo, Nguy n Thành ạt, Nguy n Quang ào (1999), M t
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126. Walker G.M., Stewart G.G (2016), Saccharomyces cerevisiae in the
Production of Fermented Beverages. Beverages, p 2, 30.
127. Walker G.M., Hill A.E (2016), Saccharomyces cerevisiae in the Production
of Whisk(e)y. Beverages 2, 38. pp 2 - 15
128. Walker G.M (2014), Microbiology of wine-making. In Encyclopedia of
Food Microbiology; Batt, C., Tortorello, M.L., Eds.; Elsevier Science
Publishers: Boston, MA, USA, 2014; pp. 787–792.
157
129. Walker G.M (2014), Fermentation (Industrial). Media for Industrial
Fermentations. In Encyclopedia of Food Microbiology; Batt, C., Tortorello,
M.L., Eds.; Elsevier Science Publishers: Boston, MA, USA, pp 162 – 198.
130. Wang Z.X., J. Zhuge H. Fang and Prior B. A (2001), Glycerol production by
microbial fermentation: A review. Biotechnology Advances. 19: 201-223.
131. Watson, DC (1981), The development of specialised yeast strains for use in
Scotch whisky fermentations. In: Current Developments in Yeast Research.
Advances in Biotechnology. Eds. GG Stewart and I Russell. Pergamon
Press, Oxford, pp 57-62.
132. Watson, D.C (1984), Distilling Yeasts, Developments in Industrial
Microbiology, 25, 213-220.
133. Watson E.B, Othman DB, Luck JM, Hofmann AW (1987), Partitioning of
U, Pb, Cs, Yb, Hf, Re and Os between chromian diopsidic pyroxene and
haplobasaltic liquid. Chemical Geology, Volume 62, Issues 3–4, Pages
191-208
134. Will Kew, Ian oodall, David larke, Dušan Uhrín (2017), hemical
Diversity and Complexity of Scotch Whisky as Revealed by High-
Resolution Mass Spectrometry. Journal of The American Society for Mass
Spectrometry January, Volume 28, Issue 1, pp 200–213.
135. Xingyan Liu, Bo Jia, Xiangyu Sun, Jingya Ai, Cheng Wang, Fang Zhao,
Jicheng Zhan, Weidong Huang (2015), Effect of Initial PH on Growth
Characteristics and Fermentation Properties of Saccharomyces cerevisiae.
Food Science. Volume80, Issue4, pp800 - 808
136. Yaya Liu, Anissa el Masoudi, Jack T. Pronk, Walter M. van Gulik (2019),
Quantitative physiology of non-energy-limited retentostat cultures of
Saccharomyces cerevisiae at near-zero specific growth rates. It is made
available under a CC-BY-NC-ND 4.0 International license
137. Yin-Won Lee1 (2011), Fungi Associated with the Traditional Starter
Cultures Used for Rice Wine in Korea, Dong-A University, J. Korean Soc.
Appl. Biol. Chem. 54(6), 933- 943.
138. Zhang W, Du G, Zhou J, Chen J (2018), Regulation of sensing,
transportation, and catabolism of nitrogen sources in Saccharomyces
cerevisiae. Microbiol Mol Biol, Volume 82 Issue 1. P 2 – 19.
158
III. 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)