甘蔗糖蜜为原料发酵合成茁霉多糖及其在芒果常温保鲜中的应用研究
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摘要
茁霉多糖(Pullulan)是出芽短梗霉利用糖发酵产生的一种胞外多糖，具有极好的成膜、成纤维、阻气、粘接、易加工、无毒性等特性，是微生物多糖中最令人瞩目的多糖之一。本文以出芽短梗霉(Aureobasidium pullulans)As3.3984作为甘蔗糖蜜发酵合成茁霉多糖的生产菌株，对甘蔗糖蜜发酵合成茁霉多糖可行性、出芽短梗霉As3.3984的生长曲线、甘蔗糖蜜合成茁霉多糖发酵条件、发酵动力学过程、多糖膜电镜结构及在芒果常温保鲜中的应用等方面进行了系统地研究。
甘蔗糖蜜含丰富的可发酵的碳源和氮源等营养成分，可以用来发酵合成茁霉多糖，但发酵条件需要优化。经甘蔗糖蜜耐受性试验表明：发酵液中甘蔗糖蜜的浓度不宜超过200g／L。测定出芽短梗霉菌As3.3984的生长曲线，认为发酵种子液的菌龄以12-18h为佳。
经比较研究发现，蒽酮法较适合茁霉多糖的检测，该方法的平均相对误差为0.6％，具有较好的准确性。
为了探讨甘蔗糖蜜为原料发酵合成茁霉多糖的初始条件，本研究以生物量和多糖产量为依据，确定以甘蔗糖蜜作为发酵合成茁霉多糖的初始发酵培养基，该发酵培养基是经预处理后的甘蔗糖蜜质量浓度为120g／L时，硫酸铵质量浓度0.4g／L，硫酸镁质量浓度1.0g／L，氯化钼质量浓度0.4g／L，发酵初始pH为5.5。研究结果表明甘蔗糖蜜为原料发酵合成茁霉多糖的生物合成条件是：接种量为15％，发酵温度为30℃，在充足供氧的条件下发酵时间为144h。
通过研究等pH法发酵技术对合成多糖的影响发现等pH发酵技术可提高多糖合成量，缩短发酵时间24h，最大合成量可到25.09g／L。
采用Logistic equation模型和Luedeking-piret方程对茁霉多糖发酵进行了较好的理论描述。通过研究建立了出芽短梗霉As3.3984生长、茁霉多糖
周文化2仪冈甘蔗糖蜜为原料发醉合成茁霉多特及其在芒果常温保鲜中的应用研究
合成和底物(糖蜜)消耗相关的动力学模型参数，阐述了茁霉多糖的动力学
特征
红外光谱表明:发酵合成的多糖的结构与茁霉多糖标准品一样。
电镜观察表明:多糖形成的网状结构膜可涂覆在芒果表面，可以封闭芒
果表面气孔。
根据测定采后芒果呼吸作用产生的C02浓度(分压)的变化，建立了
芒果呼吸动力学方程式为RcoZ=0.33lly，.9l‘2(未经涂膜处理)和Rco2==6.15
xxo‘ZlyZ·2723(经涂膜处理)。
实验表明:茁霉多糖膜在适宜的浓度下与聚乙烯薄膜协同作用对芒果外
部感官和内部品质有一定的影响，其中经3%多糖涂膜处理的芒果贮藏效果
最佳。
Pullulan,a kind of exopolysaccharides produced by Aureobasidium pullulans,has a number of favorable performances such as film-forming,high adhesion,low digestibility,nontoxicity and other characters and is used widely in food ,pharmaceutical,chemical industry.petrochemical and light industries etc. It is even drawing more and more attentions. In this paper.production of pullulan with apure culture of Aureobasidium pullulansAs3.3894 was investigated and fermentation was conducted on cane molasses,which was used as as carbon source.We also systematically studied the feasibility of fermention on cane molasses.the growth curve of As3.3894,the conditions for pullulan production on cane molasses.the dynamic processes of fermention.the SEM structure of pullulan film and the application of the film in mango fruit preservation.
Cane molasses is full of fermentable components such as carbon and nitrogen source, which can be used as produce pullulan, but the fermentation conditions should be optimiazed. The tolerable test showed that concentration of cane molasses below 200g/L was feasible for A. pullulansAs3.3984 to grow and produce pullulan. The growth curve of A. pullulansAs3.3984 indicateed that its exponential phase is 12-18h after vaccinated.
The anthranone method used as pullulan assay was studied, and this method had an average relative error of 0.6%.
The effect of molasses, nitrogen source, salt and the optimum initial pH value of medium on pullulan production of A. pullulansAs3.3984 were studied. Based on biomass and yield of polysaccharide, the suitability of fermatable medium was obtained, Compositions of the initial medium were as following:canemolasses, 120g/L;(NH4)SO40.4g/L;MgSO4,1.0g/L;MoCl2.0.4g/L, and pH,5.5.Exopolysaccharides production was studied when As3.3984 was growing in of fermatable medium and the fermenting conditions were suitable.the best culture conditions for A. pullulansAs3.3984 to synthesis pullulan were that inoculation amount is 15% of cultures ,with fermentation at
30 and adequated oxygen for 144h.
Constant pH method for exopolysaccharides produced by A. pullulansAS3.398 was studied .It was showed that the method could both increase yield of exopolysaccharides and deduce the fermentation time, and the maximum yield of pullulan was 25.09g/L and fermention time was J20h.
The pullulan production by batch fermentation of A.pullulansAs3.398 as well as the kinetics of fermention process were studied Based on Logistic equation and Luedeking-piret,the dynamic equation describing the pullulan production , biomass substrate (cane molasses ) consumption were obtained .At the same time the experimental data and the model values were tested .veified and compared.
The structure of pulluan was studied by applying Fourier transform infrared(FTIR) techniques. The FTIR spectra of pullulan showed that the structure of pullulan sample from the fermention process was the same as the standard.
the surface tructure of pullulan film was observed by applying Sanning Electron Microscopy (SEM), it showed that the surface of mango fruit was coated by the film and its widpipe was sealed.
Respiration kinetic of mango fruit before and after being coated were studied, and dynamics equations among respiratory intensity (Rco2), coefficient of penetrating(Pg), temperture(T), the carbon dioxide concentration(Y), time(t) and etc. were established .the dynamice equations were as follows: Rco2=0.331/y1.9112 (uncoated) and RCO2=6.15 10-2/y2.2723 (coated). The preserving effects to mango of pullulan film were analysed.
The experimental results showed that the sense and internal quality mango fruit were influced by pullualn film working in coordination with PE film, Excellent quality and the best presreving effect were obtained. when mango fruit was coated by 3%pullulan film
甘蔗糖蜜含丰富的可发酵的碳源和氮源等营养成分，可以用来发酵合成茁霉多糖，但发酵条件需要优化。经甘蔗糖蜜耐受性试验表明：发酵液中甘蔗糖蜜的浓度不宜超过200g／L。测定出芽短梗霉菌As3.3984的生长曲线，认为发酵种子液的菌龄以12-18h为佳。
经比较研究发现，蒽酮法较适合茁霉多糖的检测，该方法的平均相对误差为0.6％，具有较好的准确性。
为了探讨甘蔗糖蜜为原料发酵合成茁霉多糖的初始条件，本研究以生物量和多糖产量为依据，确定以甘蔗糖蜜作为发酵合成茁霉多糖的初始发酵培养基，该发酵培养基是经预处理后的甘蔗糖蜜质量浓度为120g／L时，硫酸铵质量浓度0.4g／L，硫酸镁质量浓度1.0g／L，氯化钼质量浓度0.4g／L，发酵初始pH为5.5。研究结果表明甘蔗糖蜜为原料发酵合成茁霉多糖的生物合成条件是：接种量为15％，发酵温度为30℃，在充足供氧的条件下发酵时间为144h。
通过研究等pH法发酵技术对合成多糖的影响发现等pH发酵技术可提高多糖合成量，缩短发酵时间24h，最大合成量可到25.09g／L。
采用Logistic equation模型和Luedeking-piret方程对茁霉多糖发酵进行了较好的理论描述。通过研究建立了出芽短梗霉As3.3984生长、茁霉多糖
周文化2仪冈甘蔗糖蜜为原料发醉合成茁霉多特及其在芒果常温保鲜中的应用研究
合成和底物(糖蜜)消耗相关的动力学模型参数，阐述了茁霉多糖的动力学
特征
红外光谱表明:发酵合成的多糖的结构与茁霉多糖标准品一样。
电镜观察表明:多糖形成的网状结构膜可涂覆在芒果表面，可以封闭芒
果表面气孔。
根据测定采后芒果呼吸作用产生的C02浓度(分压)的变化，建立了
芒果呼吸动力学方程式为RcoZ=0.33lly，.9l‘2(未经涂膜处理)和Rco2==6.15
xxo‘ZlyZ·2723(经涂膜处理)。
实验表明:茁霉多糖膜在适宜的浓度下与聚乙烯薄膜协同作用对芒果外
部感官和内部品质有一定的影响，其中经3%多糖涂膜处理的芒果贮藏效果
最佳。
Pullulan,a kind of exopolysaccharides produced by Aureobasidium pullulans,has a number of favorable performances such as film-forming,high adhesion,low digestibility,nontoxicity and other characters and is used widely in food ,pharmaceutical,chemical industry.petrochemical and light industries etc. It is even drawing more and more attentions. In this paper.production of pullulan with apure culture of Aureobasidium pullulansAs3.3894 was investigated and fermentation was conducted on cane molasses,which was used as as carbon source.We also systematically studied the feasibility of fermention on cane molasses.the growth curve of As3.3894,the conditions for pullulan production on cane molasses.the dynamic processes of fermention.the SEM structure of pullulan film and the application of the film in mango fruit preservation.
Cane molasses is full of fermentable components such as carbon and nitrogen source, which can be used as produce pullulan, but the fermentation conditions should be optimiazed. The tolerable test showed that concentration of cane molasses below 200g/L was feasible for A. pullulansAs3.3984 to grow and produce pullulan. The growth curve of A. pullulansAs3.3984 indicateed that its exponential phase is 12-18h after vaccinated.
The anthranone method used as pullulan assay was studied, and this method had an average relative error of 0.6%.
The effect of molasses, nitrogen source, salt and the optimum initial pH value of medium on pullulan production of A. pullulansAs3.3984 were studied. Based on biomass and yield of polysaccharide, the suitability of fermatable medium was obtained, Compositions of the initial medium were as following:canemolasses, 120g/L;(NH4)SO40.4g/L;MgSO4,1.0g/L;MoCl2.0.4g/L, and pH,5.5.Exopolysaccharides production was studied when As3.3984 was growing in of fermatable medium and the fermenting conditions were suitable.the best culture conditions for A. pullulansAs3.3984 to synthesis pullulan were that inoculation amount is 15% of cultures ,with fermentation at
30 and adequated oxygen for 144h.
Constant pH method for exopolysaccharides produced by A. pullulansAS3.398 was studied .It was showed that the method could both increase yield of exopolysaccharides and deduce the fermentation time, and the maximum yield of pullulan was 25.09g/L and fermention time was J20h.
The pullulan production by batch fermentation of A.pullulansAs3.398 as well as the kinetics of fermention process were studied Based on Logistic equation and Luedeking-piret,the dynamic equation describing the pullulan production , biomass substrate (cane molasses ) consumption were obtained .At the same time the experimental data and the model values were tested .veified and compared.
The structure of pulluan was studied by applying Fourier transform infrared(FTIR) techniques. The FTIR spectra of pullulan showed that the structure of pullulan sample from the fermention process was the same as the standard.
the surface tructure of pullulan film was observed by applying Sanning Electron Microscopy (SEM), it showed that the surface of mango fruit was coated by the film and its widpipe was sealed.
Respiration kinetic of mango fruit before and after being coated were studied, and dynamics equations among respiratory intensity (Rco2), coefficient of penetrating(Pg), temperture(T), the carbon dioxide concentration(Y), time(t) and etc. were established .the dynamice equations were as follows: Rco2=0.331/y1.9112 (uncoated) and RCO2=6.15 10-2/y2.2723 (coated). The preserving effects to mango of pullulan film were analysed.
The experimental results showed that the sense and internal quality mango fruit were influced by pullualn film working in coordination with PE film, Excellent quality and the best presreving effect were obtained. when mango fruit was coated by 3%pullulan film
引文
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