响应曲面法优化赤藓糖醇结晶工艺

为了开发一套赤藓糖醇的结晶工艺,该文研究了包括晶种添加量、溶液初始浓度、结晶时间和结晶温度在内的4个主要因素对赤藓糖醇结晶收率的影响。然后根据单因素试验的结果,采用响应曲面法整体优化了赤藓糖醇的结晶工艺参数。响应面优化结果显示,水溶液中赤藓糖醇结晶的最优操作条件为,晶种添加量1.0%,赤藓糖醇溶液初始质量浓度550 mg/mL,结晶时间3 h,结晶温度-4.5℃。上述条件下,赤藓糖醇的一次结晶率为52.78%。该文得到的模型可以用来优化赤藓糖醇在水溶液体系中的结晶过程。该模型化的工艺获得了较高的赤藓糖醇结晶收率。     

赤藓糖醇高产菌选育及发酵条件研究

为了提高赤藓糖醇的产量,对自选耐高渗酵母菌T-3-2进行紫外线与亚硝酸复合诱变处理,并采用响应曲面法优化了其发酵条件。复合诱变得到1株稳定高产突变株UN-11,其赤藓糖醇的产量达到78.3mg/mL,比T-3-2提高了38.8%;通过响应面分析建立了关键影响赤藓糖醇产量的二次多项式数学模型,得到最佳生产工艺条件为:发酵温度31℃、转速170r/min、接种量9.8%。模型预测结果产物浓度达到84.93mg/mL,验证实验结果产物浓度为85.25mg/mL。该模型对赤藓糖醇工业化生产有一定的指导意义。     

静息细胞转化赤藓糖醇生产L-赤藓酮糖条件优化

应用响应面法设计对Gluconobacter kondonii静息细胞转化合成L-赤藓酮糖进行优化。在单因素试验基础上,利用响应面分析法(RSM)对转化条件进行优化,并建立了各因素与L-赤藓酮糖之间的数学模型。结果表明,静息细胞浓度、p H值、赤藓糖醇浓度、反应时间以及温度对L-赤藓酮糖产量均有不同程度影响。响应面预测产L-赤藓酮糖的最佳工艺为静息细胞浓度1.65%、赤藓糖醇浓度97.9 g/L、温度30.2℃、p H值为5,反应时间18 h,此时L-赤藓酮糖产量为96.1 g/L,与响应面极值相吻合,转化率为98.2%,生产强度达到5.34 g/(L·h),表明优化方案达到了预期效果。     

Torulopsis sp.ERY237产赤藓糖醇工艺条件的研究

以Torulopsis sp.ERY237作为出发菌株,考察了不同碳源、氮源、无机盐类以及温度等因素对菌种产赤藓糖醇的影响,建立和优化了赤藓糖醇摇瓶发酵培养基配方、发酵工艺条件,同时研究了发酵过程中菌体生物量、pH值、产物浓度的动态变化。结果表明,菌株的最适培养基配方为(g/L):葡萄糖300,玉米浆3.5,C_([Cu~(2+)])1.5,C_([Mn~(2+)])10;适宜的培养条件为初始pH值自然,温度30℃,装液量50 mL/500 mL,转速200 r/min,在此条件下培养132 h赤藓糖醇产量达87.8 g/L,是优化前产量的1.9倍,发酵时间缩短了12 h。     

赤藓糖醇玫瑰花风味酸奶的研制

目的 优化赤藓糖醇玫瑰花风味酸奶的配方,丰富酸奶的种类.方法 以鲜牛乳为主要原料,以玫瑰花浸提液为辅料,采用赤藓糖醇代替白砂糖,制作出低热量更健康的风味酸奶.以感官评分为指标,选取玫瑰花浸提液添加量、赤藓糖醇添加量、发酵剂添加量和发酵时间4个因素,进行单因素实验,筛选出对酸奶风味影响较大的3个单因素进行三因素三水平响应...     

丛梗孢酵母产赤藓糖醇的诱变选育

利用微波-硫酸二乙酯复合诱变对产赤藓糖醇丛梗孢酵母E54进行处理,以高渗平板和摇瓶发酵为筛选方法,得到遗传稳定的诱变高产株EW29;再采用氮离子注入对EW29进行诱变处理,摇瓶发酵筛选得到诱变株EN59,其90h发酵液中赤藓糖醇产量达到55.13g/L,较EW29提高20.3%,较E54提高36.9%,遗传稳定性较好。对突变株EN59的发酵培养基进行了优化,在优化培养基葡萄糖250g/L、酵母膏5g/L、KH2PO4 0.3g/L、MnSO4 ·4H2O 0.04g/L、CuSO4 ·5H2O 0.03g/L,初始pH4的条件下,90h发酵液中赤藓糖醇平均产量达到69.00g/L以上。在优化培养基的基础上进行5L罐发酵放大实验,发酵126h赤藓糖醇产量达到71.14g/L。     

高产赤藓糖醇菌株RH-UV-L4-F9发酵条件的优化

以高产赤藓糖醇菌株RH-UV-L4-F9为研究对象,采用生物统计方法分别对该菌株的发酵培养基和培养条件进行优化。发酵培养基最佳配比为葡萄糖30%,酵母膏0.5%,脲1%,MgSO_4 0.05%;最适发酵条件为34℃,初始pH值6.0.摇床转数180r/min。最适条件下赤藓糖醇产量为157.4mg/mL。     

海藻糖与赤藓糖醇偶联发酵工艺研究

对陶瓷膜过滤后的海藻糖糖化液进行成分分析，确定主要成分及营养指标。结合成分分析结果，补加氮源进行偶联发酵并对氮源补加量进行优化，在确定的最佳氮源添加量下进行小试工艺的研究与优化。结果表明，偶联发酵最适氮源补加量为酵母浸膏、酵母浸粉各3 g/L；偶联发酵能够实现消耗葡萄糖的目的，发酵72 h，对照组及补氮组残糖分别降至0.09 g/L、0.05 g/L。此外，补充氮源提升了赤藓糖醇得率，但赤藓糖醇水平较低，仅为25.4 g/L，糖醇转化率为30.70%。在补充氮源的基础上，向底料培养基补加葡萄糖至终浓度200 g/L进行发酵，赤藓糖醇得率大幅度提升，发酵液赤藓糖醇浓度提升至104.62 g/L，糖醇转化率50.18%，较优化前分别提升了97.32 g/L,42.26%。该优化偶联发酵工艺降低了下游海藻糖分离纯化的难度，提高了海藻糖的生产效益。本研究为其他稀有糖的生产提供了新的思路。     

丛梗孢酵母发酵产赤藓糖醇条件的优化

丛梗孢酵母BH010是从蜂蜜样品中分离得到的产赤藓糖醇菌株。该实验研究了发酵培养基及发酵条件对丛梗孢酵母赤藓糖醇产量的影响。单因素实验及正交实验的结果表明,最佳发酵培养基及发酵条件为:葡萄糖含量(质量浓度)35%、酵母膏含量(质量浓度)1%、CaCl2.2H2O(质量浓度)0.2%,初始pH6.0,接种量1%,30℃摇瓶培养9d。最终赤藓糖醇产量为110.61g/L发酵液,比普通发酵条件下提高85.56%。     

耐高渗酵母产赤藓糖醇的影响因素

球拟酵母OS-194是一株单产赤藓糖醇的耐高渗酵母,该菌株高产赤藓糖醇的最佳培养基配方为葡萄糖10g/dL,酵母膏0.5g/dL,尿素0.1g/dL.最适培养条件是在摇瓶转速150r/min的条件下于35℃培养4d.在上述培养条件下,该菌株赤藓糖醇的耗糖转化率高达29.6%.磷是限制OS-194菌株高产赤藓糖醇的主要因素,当培养液中的磷质量浓度低于31.5mg/L时,赤藓糖醇的产量最高;随着磷质量浓度的升高,该菌株赤藓糖醇的产量降低,而酒精的产量和生物量却有明显升高.同时,OS-194菌株还能利用果糖、蔗糖和D-甘露糖产赤藓糖醇.     

两阶段调控葡萄糖质量浓度强化赤藓糖醇发酵的研究

该研究在5 L发酵罐水平上研究了不同初始葡萄糖质量浓度对解脂耶氏酵母（Yarrowia lipolytica）JZ-204生长和发酵产赤藓糖醇的影响。结果表明,100 g/L初始葡萄糖质量浓度有利于菌体生长,高初始葡萄糖质量浓度（300 g/L和400 g/L）有利于赤藓糖醇合成。基于此,提出两阶段葡萄糖质量浓度调控策略,即0 h时以初始葡萄糖质量浓度为100 g/L,22 h后通过补加葡萄糖使总糖量达300 g/L进行发酵。结果表明,与分批发酵相比（100 g/L、200 g/L、300 g/L、400 g/L）,采用该调控策略,赤藓糖醇产量达到最高水平92.66 g/L,分别比分批发酵提高了1 347.81%、84.54%、14.66%、7.57%;生产强度达到最高的0.48 g/（L·h）,分别比分批发酵提高了300%、37.14%、29.73%、20.00%。该调控策略为赤藓糖醇的高效发酵合成提供参考。     

Generation of an antibody specific to erythritol, a non-immunogenic food additive

Erythritol, a simple sugar alcohol, is widely used as a food and drug additive owing to its chemical inertness, sweetness and non-toxicity. Adverse reactions to erythritol are rare and only three cases of allergic reactions to foods containing erythritol have been reported. Being inert, erythritol cannot produce an immunological response. In order to explain the mechanism of immunogenicity of erythritol, a method to obtain erythritol epitopes on a carrier protein, which can serve as an immunogen to develop antibodies against erythritol, is described. D-Erythrose was conjugated to bovine serum albumin at pH 8 by reductive amination. The reduction product of the Schiff base of D-erythrose-bovine serum albumin conjugate creates erythritoyl groups. Rabbits immunized with erythritol-bovine serum albumin conjugate (29 haptens/molecule) showed good antibody response (detection of 1 µg antigen, erythritol-keyhole limpet haemocyanin conjugate possessing 50% modified amino groups, at 1 : 50 000 dilution). Anti-erythritol immunoglobulin-G antibodies were purified from the immune serum using hapten-affinity chromatography on an erythritol-keyhole limpet haemocyanin-Sepharose CL-6B affinity matrix. The yield of erythritol-specific antibody was approximately 40 µg ml^-1 of rabbit antiserum. Enzyme-linked immunobsorbant assay inhibition studies using sugars, sugar alcohols and L-lysine showed minimal cross-reactivity (approximately 4%) when compared with erythritol; only dithioerythritol showed a cross-reactivity of approximately 33%. D-Threitol and L-threitol (isomers of erythritol) had cross-reactivities of 15 and 11%, respectively. The inhibition studies confirmed the haptenic nature of erythritol and indicated that the erythritoyl group is a single epitope. The reaction scheme outlined here for the generation of erythritol epitopes appears to provide a basis for the immunogenicity of erythritol.     

Erythritol production by <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> mutant strain M53 generated through atmospheric and room temperature plasma mutagenesis

Mutants of Yarrowia lipolytica with high erythritol production were generated through an atmospheric and room temperature plasma (ARTP) mutation system. Among these mutants, Y. lipolytica M53 exhibited the highest erythritol yield. In a batch culture, M53 produced 64.8 g/L erythritol from 100 g/L glycerol. The yields of byproducts (e.g. mannitol, arabitol, and α-ketoglutaric acid) were low, and the mechanisms underlying these changes were examined by measuring enzyme activities in the pentose phosphate pathway. Up to 145.2 g/L erythritol was produced by M53 from 200 g/L of glycerol, and erythritol accumulation was promoted by 3.7 mg/L of Cu²⁺, 10.15 mg/L of Mn²⁺, and 30.37 g/L of NaCl. Fed-batch cultivation of M53 in a 5-L fermentor produced 169.3 g/L erythritol with low levels of byproducts within 168 h. This finding confirmed the potential of M53 as an erythritol producer on a commercial scale.     

无机盐渗透压对假丝酵母SK25.001发酵生产赤藓糖醇的影响

以假丝酵母SK25.001为生产菌,通过研究其发酵产赤藓糖醇的碳源、氮源、碳氮比以及NaCl、KCl对其发酵产赤藓糖醇的影响,来探索无机盐(NaCl,KCl)渗透压对赤藓糖醇发酵的影响。结果发现,葡萄糖、酵母粉分别是其最佳碳源和氮源,最佳碳氮比为20∶1,转化率达到了14.2%;向发酵培养基中添加不同浓度的KCl或NaCl后发现,菌体生长速度随着KCl或NaCl浓度增大而降低,在KCl浓度为0.4 mol/L或NaCl浓度为0.3 mol/L时赤藓糖醇产量达到最大,达到了18.4 g/L和17.4 g/L;将NaCl和KCl的浓度用渗透压表示发现赤藓糖醇的转化率随着渗透压的增大而升高,高渗透压抑制菌体的生长。     

High-level production of erythritol by strain 618A-01 isolated from pollen

We have isolated a microorganism (strain 618A-01) from pollen which has the ability to produce erythritol when grown in the presence of glucose as the carbon source. When cultivated in a medium consisting of 20% glucose and 1% dried bouillon in a shake flask, 75 g/l erythritol was produced after 950 h, corresponding to a 37.5% yield against glucose consumption. No other polyols, including glycerol, were detected in the medium. Positive-ion fast atom bombardment mass spectrometry and 1H- and 13C-NMR analyses confirmed that the fermentation product was erythritol. Scanning electron microscopic analysis clearly demonstrated that the cells grown on YPD medium at 30 degrees C showed yeast-like morphology, while they appeared like hyphae at 37 degrees C. The complete 18S rRNA sequence of the isolate was determined, which showed high identity (99.5%) with the genus Ustilago of the phylum Basidiomycota. The data strongly suggest that strain 618A-01 belongs to the class Ustilaginomycetes. The culture conditions for the production of erythritol by the isolate were examined. The use of medium containing 1% tryptone, 0.5% yeast extract and 0.5% NaCl yielded the highest cell growth and erythritol productivity among the media tested. Continuous glucose feeding at 6-7% to the fermentor further increased the production of erythritol, and we obtained a maximal 100 g/l erythritol after 530 h, with a 39.3% yield.     

利用NMR研究赤藓糖醇对糙米面包贮藏期间保水性的影响

利用低场核磁共振（nuclear magnetic resonance,NMR）及成像技术,研究赤藓糖醇和蔗糖对糙米面包贮藏期间保水性的影响。通过检测面包1H NMR弛豫时间、峰面积、核磁共振成像（magnetic resonance imaging,MRI）以及水分活度,得出贮藏期间面包结合水（弛豫时间T21）相对稳定,不易流动水（弛豫时间T22）和自由水（弛豫时间T23）逐渐减少,与蔗糖面包相比,添加赤藓糖醇的面包具有高水分含量和低水分活度的特点,MRI同样体现出添加赤藓糖醇的面包具有良好的保水性。     

Key role for transketolase activity in erythritol production by Trichosporonoides megachiliensis SN-G42

Erythritol is an important sugar alcohol industrially produced only by fermentation. The highly osmophilic yeast-like fungi, Trichosporonoides megachiliensis SN-G42, enables commercial production of erythritol with a high conversion from glucose to erythritol of more than 47%. However, the microbial production pathway of erythritol remains unclear. In the present study, the activities of enzymes in the pentose phosphate pathway of Trichosporonoides megachiliensis SN-G42 used for industrial erythritol production were measured under various culture conditions to examine the production mechanism and the key-enzymes.As a result, the various enzyme activities of this organism are revealed in the pentose phosphate pathway, i.e., those of hexokinase, glucose-6-phosphate dehydrogenase, gluconate dehydrogenase, transketolase, transaldolase, and erythrose reductase. In the cultures in which erythritol was produced after completion of cell growth, the enzyme activities of the pentose phosphate pathway were higher than those of the TCA cycle. In particular, transketolase activity was correlated with erythritol productivity under various production cultures with different agitation speeds and thiamine concentrations.These results suggest that erythritol may be produced mainly through the pentose phosphate pathway. In addition, the high activity of transketolase is required to produce abundant intermediates, which results in high erythritol productivity. As such, transketolase appears to be a key-enzyme for erythritol production in the organism studied.     

Effect of using Erythritol as a Sucrose Replacer in Making Spanish Muffins Incorporating Xanthan Gum

Spanish muffins are sweet, high-calorie baked products which are highly appreciated for their good taste and soft texture. The aim of this work was to evaluate the suitability of erythritol and of its combination with xanthan gum and double quantities of leavening agent for replacing sucrose in Spanish muffins and to understand their functionality in a muffin system. The linear viscoelastic properties of the batter during heating, its specific gravity and bubbles, muffin weight loss during baking and muffin bubbles, height, volume, and instrumental texture were studied. Both erythritol and sucrose increase in the temperature at which the viscoelastic functions increased with temperature. In comparison with the reduced sucrose muffins, the use of erythritol increased the number of air bubbles in the batter. The height of the muffins also increased in the presence of erythritol when compared to the corresponding reduced sucrose muffins, although the volume did not. Erythritol was not effective in diminishing the increased hardness associated with sucrose reduction, but the combination of erythritol with xanthan gum and a double quantity of leavening agent significantly improved the muffin volume (from 94 cm³ for 100% erythritol formulation to 108 cm³ for 100% erythritol–xanthan double-leavening agent formulation) and significantly decreased the hardness (from 75 N for 100% erythritol formulation to 25 N for 100% erythritol–xanthan double leavening agent formulation).     

碳源对圆酵母(Torula sp.)B84512发酵合成赤藓糖醇及副产物甘油的影响

研究了圆酵母(Torula sp.)B84512以不同碳源发酵产赤藓糖醇过程中副产物甘油的生成与消耗情况。发现该菌株在以任何碳源为底物发酵过程中均会产生甘油,且在发酵中后期甘油逐渐被消耗。以甘油为唯一碳源时该菌株合成赤藓糖醇的速率及产率均低于葡萄糖。葡萄糖为圆酵母B84512发酵产赤藓糖醇的最佳碳源。采用分批补料的方式提高赤藓糖醇的产率并期望能抑制甘油的生成,实验结果表明补料至总糖浓度为50%时赤藓糖醇产量最高为253 g/L,产率为1.03 g/(L.h)。但甘油产量与葡萄糖的浓度呈正相关,分批补料并不能有效抑制甘油的生成,反而导致发酵周期大大延长,对于工业化生产极其不利。通过对甘油的生成及消耗过程中关键酶胞浆3-磷酸甘油脱氢酶(ctGPD)、3-磷酸甘油酯酶(GPP)、线粒体3-磷酸甘油脱氢酶(mtGPD)酶活测定,确定胞浆3-磷酸甘油脱氢酶为甘油合成途径的关键酶,为以后对圆酵母B84512中甘油代谢途径的基因工程改造选育奠定了基础。