花粉蜂蜜酒的研制
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摘要
蜂蜜营养丰富,经济价值高,是有保健作用的甜味剂;花粉也含有多种营养物质。因此利用蜂蜜为原料,添加花粉,经酵母发酵,可生产花粉蜂蜜酒。目前国内外对酒的研究很多,但是尚未见到对花粉蜂蜜酒的研究。鉴于此,本论文对花粉蜂蜜酒的研制进行了一定研究。首先利用紫外线诱变酵母菌,对诱变后的酵母菌做了相关性能的测定,筛选出适合蜂蜜和花粉发酵用的酵母菌;对花粉破壁做了研究,并用毛细管电泳测定酒中的氨基酸的种类及含量,以此优化花粉的最佳添加方式,并计算花粉的破壁率;然后研究花粉蜂蜜酒工艺,优化酵母菌的最佳发酵条件,并测定了发酵过程中相关的理化指标;对成品酒进行了感官测评和微生物指标的检测;最后对成品酒的香气进行GC-MS测定。以期为花粉蜂蜜酒的进一步研究提供理论基础和科学依据。主要研究结果有:
1.将葡萄酒酵母经过一系列的分离纯化得到了出发菌株,对出发菌株进行不同照射时间、不同照射距离的紫外诱变处理后,获取了一定数量酵母突变体,然后经过筛选,对酵母突变体进行了酒精发酵等相关性能的测定,结果确定诱变的最佳工艺为:照射剂量为15 w、照射距离为20 cm、照射时间为60 s。
2.运用毛细管电泳法来测定成品酒中的氨基酸,分析优化出最佳的花粉破壁方法:于发酵旺盛期添加发酵液的0.5%的温差法破壁的油菜花粉,经过10天主发酵期后,可以获得氨基酸含量高,品质良好的花粉蜂蜜酒。
3.本实验分别采用超声破碎法、酶法和温差破壁法这几种不同方式处理花粉,用血球计数板计算其破壁率,并用光学显微镜观测花粉破壁形态,发现温差法破壁的花粉破壁率最高,高达74.3%。
4.通过对花粉蜂蜜酒的工艺进行研究,其最佳发酵条件为:即用蜂蜜为原料,加水稀释调整其糖度为20%,添加0.25g/L的氯化镁、0.25g/L的氯化钙的营养盐和90mg/L的SO2;当诱变的液态酵母种子的接种量为9%,发酵温度为25℃,在发酵旺盛期添加0.5%温差法破壁的油菜花粉,经过10天主发酵期后,可以获得品质良好的花粉蜂蜜酒。
5.采用CHCl2萃取花粉蜂蜜酒中的香气成分,经GC-MS分析测定,分离出12个峰,鉴定出7种化合物。香气物质以醇类和脂类为主,包括苯乙醇、己酸乙酯、n-癸酸、4-羟基苯乙醇、3,7,11-三甲基-2,6,10-十二碳三烯-1-醇、7-甲氧基-2,2,4,8-四甲基三环十一烷和1,2-苯二羟酸-2-甲基丙酯这几种香气物质。
Honey with abundant of nutritious value and high economical value, is a sweetening agent with health-care function. Pollen also has full of nutrients. So pollen mead can be obtained by yeast’s fermenting the mixture of honey and pollen, with honey as the main ingredient. Now there are many studies on wine both in and out of China, but we haven’t seen a report on pollen mead. So this paper was made a study on it. First the yeast was mutated by ultraviolet ray, and the pertinent capability of yeast was measured. A research on breaking the pollen wall was made and a suitable method for breaking the pollen wall was found, and the rate of wall-broken was counted. Then a study on the fermentation of pollen mead was made and the optimum fermentation conditions were obtained. Capillary electrophoresis chromatography was used to analyze the kinds and contents of amino acids in the mead and to find a suitable method for breaking the pollen wall, and the relative physical and chemical marks of pollen mead in the process of fermentation were surveyed. The sensory evaluation and microbial marks of pollen mead were determined. Finally the aroma components of mead were analyzed by GC-MS. All the experiments were provided in a view to theoretical foundation and scientific basis for further study of pollen mead.
The results are as follows:
1. A initial strain was got by a series of purification of wine yeast. Several numbers of mutant yeast strains, obtained from the initial strain by UV irradiating for different time and different distance, were screened. Then a survey on the pertinent capability of mutant strain was determined. The results showed that the optimum mutant conditions were the irradiating dosage of 15 w, the irradiating distance of 20 cm and the irradiating time of 60 s.
2. Capillary electrophoresis chromatography was used to analyze the kinds and contents of amino acids in the pollen mead and to analyze a suitable method for breaking the pollen wall. In the period of active fermentation, 0.5% rape pollen whose wall was broken by the temperature fluctuation was added into the fermented honey. After 10 days of the period of main fermentation, the pollen mead produced in this way was rich in amino acids with good quality.
3. This test used several different methods for breaking the rape pollen wall: ultrasonic method, enzyme based method, and temperature fluctuation. The haemacytometer was used to count the rate of wall-breaking of pollen, and the morphologic changes of the wall-broken pollen were observed under the light microscope. We found that temperature fluctuation had the highest wall-breaking rate of pollen, and the rate was high to 74.3%.
4. In order to get the optimum fermenting conditions, a study on pollen mead was made. The results showed that: with honey as the main ingredient, adding water into the honey to adjust the initial sugar concentration of 20%, and adding the nutrient salts of 0.25g/L MgCl2 and 0.25g/L CaCl2 , the SO2 addition was 90 mg/ L, the inoculation of yeast was 9%, the fermenting temperature was 25℃, in the period of growing abundantly and lively of the fermentation, 0.5% rape pollen whose wall was broken by the temperature fluctuation was added into the fermented honey, after 10 days of the period of main fermentation, the pollen mead with high quality can be obtained.
5. Aroma components in the pollen mead were extracted by solvent extraction. According to the analysis of gas chromatography/mass spectrometry, 12 peaks were separated and 7 components were identified. The main components were phenethyl alcohol, ethyl caproate, n-decanoic acid, 4-hydroxy phenethyl alcohol, 3,7,11-Trimethyl-2,6,10-dodecatrien-1-ol, 7-anisyl-2,2,4,8-tetramethyltricyclo undecane, 1,2-Benzenedicarboxylic acid-2-Methyl- propyl ester.
1.将葡萄酒酵母经过一系列的分离纯化得到了出发菌株,对出发菌株进行不同照射时间、不同照射距离的紫外诱变处理后,获取了一定数量酵母突变体,然后经过筛选,对酵母突变体进行了酒精发酵等相关性能的测定,结果确定诱变的最佳工艺为:照射剂量为15 w、照射距离为20 cm、照射时间为60 s。
2.运用毛细管电泳法来测定成品酒中的氨基酸,分析优化出最佳的花粉破壁方法:于发酵旺盛期添加发酵液的0.5%的温差法破壁的油菜花粉,经过10天主发酵期后,可以获得氨基酸含量高,品质良好的花粉蜂蜜酒。
3.本实验分别采用超声破碎法、酶法和温差破壁法这几种不同方式处理花粉,用血球计数板计算其破壁率,并用光学显微镜观测花粉破壁形态,发现温差法破壁的花粉破壁率最高,高达74.3%。
4.通过对花粉蜂蜜酒的工艺进行研究,其最佳发酵条件为:即用蜂蜜为原料,加水稀释调整其糖度为20%,添加0.25g/L的氯化镁、0.25g/L的氯化钙的营养盐和90mg/L的SO2;当诱变的液态酵母种子的接种量为9%,发酵温度为25℃,在发酵旺盛期添加0.5%温差法破壁的油菜花粉,经过10天主发酵期后,可以获得品质良好的花粉蜂蜜酒。
5.采用CHCl2萃取花粉蜂蜜酒中的香气成分,经GC-MS分析测定,分离出12个峰,鉴定出7种化合物。香气物质以醇类和脂类为主,包括苯乙醇、己酸乙酯、n-癸酸、4-羟基苯乙醇、3,7,11-三甲基-2,6,10-十二碳三烯-1-醇、7-甲氧基-2,2,4,8-四甲基三环十一烷和1,2-苯二羟酸-2-甲基丙酯这几种香气物质。
Honey with abundant of nutritious value and high economical value, is a sweetening agent with health-care function. Pollen also has full of nutrients. So pollen mead can be obtained by yeast’s fermenting the mixture of honey and pollen, with honey as the main ingredient. Now there are many studies on wine both in and out of China, but we haven’t seen a report on pollen mead. So this paper was made a study on it. First the yeast was mutated by ultraviolet ray, and the pertinent capability of yeast was measured. A research on breaking the pollen wall was made and a suitable method for breaking the pollen wall was found, and the rate of wall-broken was counted. Then a study on the fermentation of pollen mead was made and the optimum fermentation conditions were obtained. Capillary electrophoresis chromatography was used to analyze the kinds and contents of amino acids in the mead and to find a suitable method for breaking the pollen wall, and the relative physical and chemical marks of pollen mead in the process of fermentation were surveyed. The sensory evaluation and microbial marks of pollen mead were determined. Finally the aroma components of mead were analyzed by GC-MS. All the experiments were provided in a view to theoretical foundation and scientific basis for further study of pollen mead.
The results are as follows:
1. A initial strain was got by a series of purification of wine yeast. Several numbers of mutant yeast strains, obtained from the initial strain by UV irradiating for different time and different distance, were screened. Then a survey on the pertinent capability of mutant strain was determined. The results showed that the optimum mutant conditions were the irradiating dosage of 15 w, the irradiating distance of 20 cm and the irradiating time of 60 s.
2. Capillary electrophoresis chromatography was used to analyze the kinds and contents of amino acids in the pollen mead and to analyze a suitable method for breaking the pollen wall. In the period of active fermentation, 0.5% rape pollen whose wall was broken by the temperature fluctuation was added into the fermented honey. After 10 days of the period of main fermentation, the pollen mead produced in this way was rich in amino acids with good quality.
3. This test used several different methods for breaking the rape pollen wall: ultrasonic method, enzyme based method, and temperature fluctuation. The haemacytometer was used to count the rate of wall-breaking of pollen, and the morphologic changes of the wall-broken pollen were observed under the light microscope. We found that temperature fluctuation had the highest wall-breaking rate of pollen, and the rate was high to 74.3%.
4. In order to get the optimum fermenting conditions, a study on pollen mead was made. The results showed that: with honey as the main ingredient, adding water into the honey to adjust the initial sugar concentration of 20%, and adding the nutrient salts of 0.25g/L MgCl2 and 0.25g/L CaCl2 , the SO2 addition was 90 mg/ L, the inoculation of yeast was 9%, the fermenting temperature was 25℃, in the period of growing abundantly and lively of the fermentation, 0.5% rape pollen whose wall was broken by the temperature fluctuation was added into the fermented honey, after 10 days of the period of main fermentation, the pollen mead with high quality can be obtained.
5. Aroma components in the pollen mead were extracted by solvent extraction. According to the analysis of gas chromatography/mass spectrometry, 12 peaks were separated and 7 components were identified. The main components were phenethyl alcohol, ethyl caproate, n-decanoic acid, 4-hydroxy phenethyl alcohol, 3,7,11-Trimethyl-2,6,10-dodecatrien-1-ol, 7-anisyl-2,2,4,8-tetramethyltricyclo undecane, 1,2-Benzenedicarboxylic acid-2-Methyl- propyl ester.
引文
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