啤酒酿造中双乙酰的微生物代谢工程调控

双乙酰是啤酒中的重要风味物质，也是影响啤酒成熟和质量的关键因素之一。以酵母细胞内双乙酰的代谢途径和微生物代谢工程为基础，通过分析细胞内双乙酰的代谢网络，设计合理的重组DNA——增加ILV5和ILV3基因拷贝数，提高缬氨酸生产通量；改变ILV2基因，降低双乙酰的前驱物质α-乙酰乳酸的生成；构建含α-乙酰乳酸脱羧酶（ALDC）基因的工程菌；在啤酒发酵过程中外加α-乙酰乳酸脱羧酶制剂——以此来降低或阻断双乙酰的生成，从而缩短啤酒的发酵周期。本文综述了啤酒酿造中双乙酰的微生物代谢工程调控的研究进展。     

啤酒酿造中的双乙酰及其酶法控制研究进展

啤酒成熟的一个重要方面是啤酒风味物质的成熟,双乙酰是主要指标之一.双乙酰含量是成熟期的时间限制因子,双乙酰在啤酒罐装后的回升问题也不容忽视.啤酒酿造中控制双乙酰的一个主要方法是以α-乙酰乳酸脱羧酶来改变代谢支路,将双乙酰的前体物质α-乙酰乳酸直接快速地转化为乙偶姻,从而降低α-乙酰乳酸转变为双乙酰的量,缩短啤酒成熟时间,减少成品啤酒中双乙酰的回升,提高啤酒风味稳定性.本文主要介绍双乙酰的代谢途径、α-乙酰乳酸脱羧酶在啤酒酿造中降低双乙酰的作用机理以及国内外有关α-乙酰乳酸脱羧酶的研究进展情况.     

生物技术在控制啤酒中双乙酰含量方面的应用

双乙酰是啤酒中的重要风味物质，其含量是控制啤酒质量的一个重要指标。近年来许多生物技术广泛应用于啤酒生产，发酵过程中加入α-乙酰乳酸脱羧酶，可降低双乙酰的峰值和在酒液中的含量，缩短发酵时间。最适pH为6，最适温度为35～40℃。磁性固定化ALDC的应用可简单方便地与酒液分离，不影响啤酒风味，从而实现生产的连续化、自动化，缩短生产周期，大大降低生产成本。LLS催代酶的应用可促进双乙酰的还原、减少α-乙酰乳酸含量，防止双乙酰反弹。构建酵母工程菌株和选育低双乙酰产生的酵母菌，可构建低乙酰乳酸合成酶，催化丙酮酸和活性乙醛在胞内过量合成双乙酰的前体物质乙酰乳酸．从而降低双乙酰含量。(孙悟)     

啤酒中的双乙酰

双乙酰是啤酒中的风味物质，主要由前体物质α－乙酰乳酸在酵母细胞外经非酶氧化脱羧作用形成，然后在细胞内由酵母还原酶作用而消除。控制双乙酰含量的途径：减少α－乙酰乳酸的生成；加速α－乙酰乳酸的分解和双乙酰的还原。具体鬼话为：强化酵母质量管理；保证麦汁组成合理；保证发酵前期的发酵速度；保证后发酵的发酵速度；双乙酰还原阶段提高罐压；增强啤酒还原性。     

啤酒发酵中双乙酰的产生和α-乙酰乳酸脱羧酶的应用

双乙酰是酵母在进行缬氨酸的合成代谢过程中产生的中间产物α-乙酰乳酸积累并分泌到酵母细胞外,通过氧化脱羧反应而产生。啤酒发酵中控制双乙酰的含量需从加快双乙酰的还原和减少α-乙酰乳酸的积累两方面综合控制。啤酒发酵中添加a-乙酰乳酸脱羧酶可将a-乙酰乳酸快速催化分解为乙偶姻,不形成双乙酰而加快啤酒成熟,显著缩短啤酒后熟发酵期。     

国产α—乙酰乳酸脱羟酶在啤酒生产中的应用

双乙酰的前驱物质是α-乙酰乳酸,在生产过程中添加α-乙酰乳酸脱羧酶可迅速降解α-乙酰乳酸,从而降低双乙酰含量,缩短发酵周期。通过两年来对进口和国产α-乙酰乳酸脱羧酶的试用比较,认为国产α-乙酰乳酸脱羧酶的使用效果完全可靠,能够替代进口同类酶制剂。     

应用包埋α-乙酰乳酸脱羧酶改进啤酒发酵及控制的模型研究

α-乙酰乳酸脱羧酶的应用提高了啤酒的发酵和成熟速度,因为它可以抑制发酵过程中双乙酰的形成.为了降低外源酶的投入成本,Claire Dulieu等设计和研究了一个含有包埋α-乙酰乳酸脱羧酶的啤酒发酵工艺及模型.就该模型研究、α-乙酰乳酸和双乙酰在啤酒发酵过程中的分布规律加以说明.     

温度对Zymomonas mobilis糖代谢关键酶活力和代谢流量的影响

以运动发酵单胞菌(Zynwmonas mobilis)ATCC31821为模式菌株,研究不同温度条件对其葡萄糖代谢关键酶活力的影响.采用全自动发酵罐,在整个发酵过程中通过充入氮气调节发酵液的溶氧量(DO)=0%,添加0.5mol/LNaOH溶液控制pH=5.5,发酵温度分别控制为25、30、35、40℃,发酵24h,测定其糖代谢网络中ED、HMP、TCA等途径的关键酶活力和代谢物成分.结果表明,在发酵温度为30～35℃时,乙醇脱氢酶(ADH)、葡萄糖-6-磷酸脱氢酶(G-6-PDH)、丙酮酸脱羧酶(PDC)、葡萄糖激酶(GK)、丙酮酸激酶(PK)和甘油醛-3-磷酸脱氢酶(GD-3-PDH)的活力较高,菌体的ED途径代谢活跃,碳素流量增加,乙醇生产量和糖转化率较高,而TCA途径的苹果酸脱氢酶(MDH)和异柠檬酸脱氢酶(ICDH)等活力较低,进入TCA途径的碳素流量明显减少;发酵温度为25、40℃时,TCA途径的酶活力升高,ED途径的酶活力减弱,生成乙醇的代谢流量减少,因此温度是z.mobilis发酵过程中调控菌体细胞生长和糖代谢的一个重要因素. 、葡萄糖-6-磷酸脱氢酶(G-6-PDH)、丙酮酸脱羧酶(PDC) 葡萄糖激酶(GK)、丙酮酸激酶(PK)和甘油醛-3-磷酸脱氢酶(GD-3-PDH)的活力较高,菌体的ED途径代谢活跃,碳素流量增加,乙醇生产量和糖转化率较高,而TCA途径的苹果酸脱氢酶(MDH)和异柠檬酸脱氢酶(ICDH)等活力较低,进入TCA途径的碳素流量明显减少;发酵温度为25、40℃时,TCA途径的酶活力升高,ED途径的酶活力减弱,生成乙醇的代谢流量减少,因此温度是z.mobilis发酵过程中调控菌体细胞生长和糖代谢的一个重要因素.葡萄糖-6-磷酸脱氢酶(G-6-PDH)、丙酮酸脱羧酶(PDC) 葡萄糖激酶(GK)、     

α-乙酰乳酸脱羧酶和α-氨基氮影响啤酒发酵中双乙酰产生的试验研究

通过对α-乙酰乳酸脱羧酶的加入与否，以及麦汁中α-氨基氮含量不同时对双乙酰产生情况的研究，结果表明：在发酵第2～12d的过程中，双乙酰变化呈现一定的规律性，在4d时达到峰值，同时α-乙酰乳酸脱羧酶的加入和高α-氨基氮含量的麦汁会减少双乙酰的产生量。     

酶共催化降低啤酒中双乙酰含量的研究设想

对啤酒生产工业中双乙酰的危害、生成机制和目前除去双乙酰的方法做了简要介绍。从实际角度出发，建设性提出了α－乙酰乳酸脱氢酶和葡萄糖氧化本 催化除去双乙酰的构想，并结合酶制剂本身性质和啤酒发酵工艺过程提出了这一构想的实现可能性。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the