The comparative ability of four isolates of Bacillus subtilis to ferment soybeans into dawadawa

In an attempt to develop starter cultures for fermenting soybeans into the traditional West African condiment dawadawa, four isolates of Bacillus subtilis: 24BP(2), 72RP(17), 72BP(30), and FpdBP(2), which had been selected from 42 Bacillus cultures in a previous study by the current authors, were used separately to produce soy-dawadawa. The accompanying microbiological and biochemical changes, including enzymatic activities, as well as the organoleptic quality of the products were evaluated including that of a control sample which was fermented spontaneously. Significant differences existed in the ability of the four isolates to hydrolyse the soybean proteins, starch, and fat to produce dawadawa. Bacillus subtilis 24BP(2) recorded the highest protease and amylolytic activities, 101 U/ml and 26.68 mg/ml, respectively, and liberated the most amino acids, 117.64 mg/g dry wt., during fermentation. Bacillus subtilis 24BP(2) also grew to the highest population of cells in the final product. Taste panelists found soybean dawadawa produced by each of the four isolates acceptable and rated soup flavoured with soy-dawadawa produced by Bacillus subtilis FpdBP(2) as the best sample. Panelists scored it higher than the control sample and soy-dawadawa produced by Bacillus subtilis 24BP(2) in that order.     

Flavouring composition prepared by fermentation with Bacillus spp

Fermented foodstuffs and condiments remain a key constituents of diets throughout many parts of Asia and Africa. In cases where the process of fermentation evolved for the development of taste or aroma, it often resulted in enhanced nutrition, stabilisation of the original raw materials, and detoxification of anti-nutrient factors. Several fermented products rely on the participation of various Bacillus species, including Bacillus natto and B. subtilis. Often, the finished products are of a very local character and exhibit sensory properties resulting from unique flora and processing technologies applied in small scale, home-based fermentations. Fermentation with B. natto and B. subtilis can produce very characteristic aromas in fermented products such as natto and dawadawa (also referred to as daddawa). Moreover, the hydrolytic capabilities of these microorganisms can result in a precursor-rich environment, which is useful for subsequent reactions leading to flavour production. A 1995 patented process demonstrated the ability to produce a fermented flavouring composition with the use of Bacillus spp. Hydrolysed protein obtained after fermentation with Bacillus spp. is mixed with reactive flavour precursors, which are subsequently heated to induce flavour formation and can be dried to a powder format. The product of this patented process imparts a basic meaty flavour, with a reduced yet characteristic dawadawa-like aroma. This paper briefly summarises some of the characteristics and uses of traditional dawadawa and illustrates alternatives described in the patent for the production of a process flavour base. Issues and considerations for the industrialisation of a fermentation process are briefly discussed, as well as some future opportunities for development and exploitation of traditional fermentation technology.     

Effect of boiling and roasting on the fermentation of soybeans into dawadawa (soy-dawadawa)

Soybeans which had initially been dehulled by either boiling (boiled/dehulled) or roasting (roasted/dehulled) before peeling, were cooked and fermented into dawadawa, a traditional food condiment. The micropopulation, enzymatic activities, proximate composition, amino acid, and aroma profiles of the two types of soybean dawadawa were evaluated during fermentation. Only minor differences were found in the microbial profiles of the two types of soy-dawadawa. Although boiled/dehulled soy-dawadawa initially had lower microbial counts, it recorded higher counts at the advanced stages of fermentation. Proteolytic and amylolytic Bacillus species including Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, Bacillus cereus, and Bacillus firmus dominated the micropopulation of the two types of soy-dawadawa with Bacillus subtilis accounting for about 50% of the Bacillus species in all samples. Lactic acid bacteria and yeasts occurred in low numbers in the two types of soy-dawadawa. The proximate composition of the two types of soy-dawadawa were similar, and their contents of moisture and protein increased whilst fat and ash decreased during fermentation. Both types of fermenting soy-dawadawa recorded similar levels of alpha-amylase activity, but boiled/dehulled soy-dawadawa showed slightly higher protease activity. The levels of isoleucine, leucine, lysine, phenylalanine, arginine and proline increased significantly with fermentation time in both types of soy-dawadawa. With respect to differences in their aroma profiles, hexanodecanol, octadecyl acetate, 1,2-dimethyl benzene, tetradecene, (E)-5-eicosene, cyclohexadecane, and hexacosane were found only in the roasted/dehulled samples, whilst 1,2-ethanediol, ethyl acetate, dimethyl disulfide, cyclotetradecane, decene, indole , 2 butyl-octenal, acetophenone, and toluene were found only in the boiled/dehulled samples. A market focus group showed preference for roasted/dehulled soy-dawadawa over boiled/dehulled soy-dawadawa. Apart from the volatile aroma compounds, the biochemical and microbiological profiles of the two types of soy-dawadawa showed only minor differences and were also similar to the profiles reported for African locust bean dawadawa.     

Identification of proteolytic bacteria from thai traditional fermented foods and their allergenic reducing potentials

ABSTRACT:  This study aimed to identify proteolytic bacteria from Thai traditional fermented foods and investigate their allergenic reducing potentials to wheat and milk allergens. Nine bacteria were isolated from fermented foods as follows: fermented soybean seeds (Thua Nao), fermented soybean paste (Thua Nao), wheat flour dough of steamed stuffed bun (Sa La Pao), and soaked rice from Thai fermented rice-noodle (Kha Nhom Jeen) processing. Both phenotypic and genotypic identifications were used in this study. It was found that all isolates were Gram-positive rods. Seven isolates were matched and identified as Bacillus subtilis by both techniques, and the remaining 2 isolates were phenotypically and genotypically identified as B. licheniformis and B. subtilis , respectively. The concentrated crude enzyme of B. subtilis DB and SR could reduce allergenicity of gliadin by hydrolyzing the allergenic gliadin fragments detected by immunoblotting. Furthermore, the enzyme of B. subtilis DB could also reduce allergenicity of β-lactoglobulin (β-LG) detected by hydrolyzing the major allergenic epitope of β-LG at Gln³⁵-Ser³⁶ position. B. subtilis DB and SR can be applied for the production of hypoallergenic wheat flour or milk food products.     

复合菌发酵腊豆的工艺优化

腊豆是中国传统的发酵豆制品,味道咸鲜适中,营养丰富,但目前腊豆大多为家庭生产且为自然发酵,存在保存时间短、口感较差等缺点,因此本实验采用Design Expert 8.06软件对枯草芽孢杆菌(Bacillus subtilis)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和酿酒酵母(Saccharomyces cerevisiae)这3株待用菌株进行复配。结果表明,在pH这个指标上,接菌组显著低于CK组,其中第6组pH显著低于其他组;接菌组质构指标显著优于CK组。在氨基态氮的指标方面,接菌组优于CK组,从游离氨基酸含量来看,第6组其鲜味氨基酸和甜味氨基酸都高于其他组,通过感官分析,第6组得分最高。综合考虑,选择第6组为最佳配比,复合菌液的配方为枯草芽孢杆菌:解淀粉芽孢杆菌:酿酒酵母为5.33:3.67:1。本实验希望通过复合菌发酵改良腊豆的发酵工艺,改善腊豆的营养等指标,促进居民营养健康状况改善。     

含水量对不同原料发酵参数的影响

试验旨在研究含水量对不同饲料原料发酵品质的影响,采用分组对照实验设计,以乳酸菌、酵母菌、枯草芽孢杆菌分别固态发酵玉米、豆粕和棉籽粕,设30%、40%、50%、60%四个含水量,以感官评价、pH和发酵后益生存留量为发酵品质评定指标。结果表明：随发酵时间延长,pH表现二次曲线的规律（P<0.01）;含水量显著影响玉米、豆粕和棉籽粕的发酵（P<0.05）,不影响原料初始pH（P>0.05）;pH达到稳定时,50%含水量的玉米、豆粕和棉籽粕的乳酸菌、枯草芽孢杆菌数量高于其它处理（P<0.05）。本试验条件下,乳酸菌、酵母菌、枯草芽孢杆菌分别发酵玉米,最佳含水量为50%;乳酸菌、枯草芽孢杆菌分别发酵豆粕和棉籽粕,最佳含水量为60%;酵母菌发酵豆粕和棉籽粕,最佳含水量为50%。     

Quality and Functional Characteristics of Chungkukjang Prepared with Various Bacillus sp. Isolated from Traditional Chungkukjang

ABSTRACT: Bacillus circulans, Brevibacillus brevis, B. licheniformis, B. coagulans, B. subtilis , and B. sterothermophillus were isolated and identified from chungkukjangs (Korean traditional soybean paste fermented for a few day). Chungkukjang was prepared on a laboratory scale with soybeans and the isolated strains. Characteristics of the chungkukjangs including slime material content, free amino acid content, sensory qualities, and antimutagenicity were determined. The content of slime material, which is an important indicator of the quality of chungkukjang, was highest in B. licheniformis -inoculated chungkukjang, andlowestin B. sterothermophillus -inoculated chungkukjang. The total content of glycine, glutamic acid, aspartic acid, and arginine, which contribute a savory taste to chungkukjangs, was highest in B. licheniformis-inoculated chungkukjang. The content of leucine, which gives a bitter taste, was highest in B. brevis -inoculated chungkukjang. Sensory evaluation revealed that chungkukjangs made using B. licheniformis and B. subtilishad a weak bitter taste and strong sweet and savory taste and good color, so their overall acceptability was high. Chungkukjang fermented with B. circulans and B. licheniformis inhibited N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) mutagenicity by more than 80%. B. licheniformis-inoculated chungkukjang exhibited the highest antimutagenicity against and aflatoxin B₁ (AFB₁) and MNNG. These results suggest that using B. licheniformis to ferment chungkukjang increases the antimutagenic properties and improves the sweet and savory taste by increasing glycine, glutamic acid, aspartic acid, and arginine concentrations.     

响应面法优化蛋壳豆渣发酵工艺生产高钙饲料

大量废弃的蛋壳和湿豆渣是良好的钙和氮素资源,为探索一条解决两类农业废弃物合理利用的新途径,本实验选用枯草芽孢杆菌、植物乳杆菌和粪肠球菌固体生料发酵鸡蛋壳、豆渣混合物。以乳酸钙含量为指标,利用单因素实验研究了豆渣蛋壳比例、固水比、葡萄糖添加量、接种量、混合菌种比例、发酵温度和发酵时间对发酵产乳酸钙的影响,并利用响应面试验优化发酵产乳酸钙的工艺条件。实验结果表明:在豆渣蛋壳比例90:10、枯草芽孢杆菌:植物乳杆菌:粪肠球菌为1:1:1(体积比)、固水比1:2.5、葡萄糖添加量15.6%、接菌量14.7%、发酵温度37℃、发酵6 d的条件下,蛋壳豆渣混合物中的乳酸钙含量达到13.58%。与发酵前相比,发酵后豆渣饲料中的游离氨基酸、乳酸钙含量显著提高(P<0.05),pH、粗脂肪、可溶性蛋白显著降低(P<0.05)。综上所述,利用益生菌发酵蛋壳与豆渣,可以制备一种高钙饲料。     

酱醅与豆酱微生物关系研究

为探究酱醅与豆酱微生物之间的关系,以两份自然发酵酱醅和一份工业发酵酱醅为对象,应用Miseq测序对酱醅及其发酵豆酱进行微生物测序分析。结果表明:在门水平上,酱醅和豆酱的优势菌门都为子囊菌门(Ascomycota)和厚壁菌门(Firmicutes);在属水平上,酱醅与豆酱中的优势真菌都为青霉菌(Penicillium)和毛霉菌(Mucor),但优势细菌的组成不同,其中两份酱醅样品(LK和SK)的优势菌属为芽孢杆菌(Bacillus)和乳杆菌(Lactobacillus),另一份酱醅样品(FK)的优势菌属为乳杆菌(Lactobacillus)、枝芽孢杆菌(Bacillus subtilis)和短乳杆菌(Lactobacillus brevis);在后期发酵过程中,虽然不同样品发酵前期细菌的群落组成不同,但随着发酵的进行,优势菌都为四联球菌(Tetragenococcus)。本研究阐明了酱醅与豆酱之间的微生物关系,为豆酱发酵过程控制提供了理论基础。     

沉香型酒醅中产香芽孢杆菌的分离鉴定及代谢产物分析

从沉香型酒醅中分离产香芽孢杆菌并进行发酵风味分析。通过平板分离得到4株产香较好的芽孢杆菌,并进行16S rDNA序列分析和构建系统发育树,4株菌分别为阿式芽孢杆菌（Bacillus aryabhattai）、地衣芽孢杆菌（Bacillus licheniform）、枯草芽孢杆菌（Bacillus subtilis）、解淀粉芽孢杆菌（Bacillus amyloliquefaciens）。其中地衣芽孢杆菌产蛋白酶和淀粉酶能力均为最强且最适生长温度为50 ℃。以玉米粉为底物进行单菌液态发酵,采用固相萃取和气相色谱-质谱法(SPME-GC-MS)分析发酵液中的风味物质,一共检测到24种风味物质,其优势产物主要为3-羟基-2-丁酮、2,3-丁二醇和一些酸类等风味物质。由此而见,阿式芽孢杆菌、地衣芽孢杆菌、枯草芽孢杆菌、解淀粉芽孢杆菌这4类菌对沉香型白酒风味的形成均具有重要作用。     

纳豆菌的分离、鉴定及产蛋白酶液态发酵条件的优化

利用奶粉豆粕培养基,采用稀释平板分离法,以H/C(透明圈直径/菌落直径)为指标,从日本传统食品纳豆中分离出9株蛋白酶产生菌。将这些初筛菌株进行发酵培养,采用Folin-酚法测定酵上清液的蛋白酶活性,选出蛋白酶活性最高的菌株H2,并依据形态特点和主要生理生化特征,鉴定为芽孢杆菌纳豆亚种(Bacillussubtilis sub.Natto)。最后以蛋白酶活性为指标,考察培养基初始pH值、温度、装液量、接种量和种龄等因素对H2菌株液态发酵的影响。结果表明,该菌株液态发酵产蛋白酶的适宜条件是:培养基初始pH值7、培养温度40℃、装液量40 mL/250 mL、接种量3%、种龄25 h。     

两步固态发酵法酿造功能性豆豉

运用分离自云南传统发酵豆豉的Bacillus subtilis SP-8-5与Lactobacillus plantarum YM-5-2菌株对大豆进行两步固态混合发酵。首先,将菌株B.subtilis SP-8-5接种至经室温浸泡10 h(20℃,质量比为1∶3),110℃(20min)蒸煮处理并冷却至30℃的大豆中,于26℃进行初步发酵42 h;随后再接种L.plantarum YM-5-2,并于30℃发酵2 d,最后置于4℃进行后发酵。经两步固态混合发酵处理后,得到一种纤溶活性较强,生产周期短,保质期相对较长,风味独特且易于被大众接受的新型功能性发酵豆豉。当后发酵时间为21 d时,样品豆豉中B.subtilisSP-8-5活菌数为(5.88±0.53)×108 CFU/g,而L.plantarum YM-5-2活菌数则高达(3.50±0.35)×1011 CFU/g,此时检测豆豉纤溶酶的纤溶圈直径高达(2.99±0.06)cm(37℃,静置培养48 h)。     

Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments

The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.     

Evaluation of different systems for the identification of Bacillus strains isolated from Spanish fermented sausages

Sixty-nine isolates obtained during the manufacture and ripening of Spanish fermented sausages were identified to species level using the Vitek Bacillus biochemical card, the dichotomous key and table proposed by Berkeley et al. ((1984). In Methods in Microbiology, Vol. 16. Academic Press, London, p. 291), morphological and physiological tests and the API 20E miniaturized system. None of the tested systems was entirely satisfactory and the final identification was mainly done on the basis of cellular morphology and the table of test results. Our isolates belonged to the species: B. subtilis (37), B. megaterium (22), B. pumilus (5), B. circulans (3) and unidentified (2). Forty-five cultures (65.2%) were accurately identified with the dichotomous key. A similar figure for the Vitek Bacillus biochemical card was 36%. The results of the API 20E system were very reproducible, especially those of the Voges-Proskauer test. Most of the strains of B. megaterium were misidentified as B. subtilis with the dichotomous key. On the other hand, a high percentage of the cultures belonging to B. subtilis were misidentified as B. megaterium with the Vitek system.     

STORABILITY AND POTABILITY OF PASTEURIZED AND STERILIZED "KUNUN-ZAKI": A FERMENTED SORGHUM BEVERAGE

The possibility of extending the shelf-life of "Kunun-zaki", a nonalcoholic sorghum based fermented beverage after pasteurization and sterilization was investigated. The traditional method of production was adopted and microbiological studies carried out. The predominant microorganisms isolated from the fermented beverage were Lactabacillus plantarum and Bacillus subtilis. Enterobacter spp. and Saccharomyces spp. occurred in small numbers. Proximate analysis of unpasteurized samples showed a decrease in total sugar and increased moisture content after one week storage at refrigeration temperature. Pasteurized samples were most acceptable to the taste panel up to two weeks of refrigerated storage, but rejected thereafter. Although the sterilized "Kunun-zaki" was shelf-stable for the two weeks of investigation, its flavor and color were adversely affected, and therefore unacceptable to the panelists. This study has shown that the shelf-life of "Kunun-zaki" (usually 24–36h) can conveniently be prolonged without serious effects on the usual organoleptic properties for up to two weeks with pasteurization.     

Quality, antioxidative ability, and cell proliferation-enhancing activity of fermented black soybean broths with various supplemental culture medium

The fermented soybean-based foods have played an important role in traditional diets around the world for many centuries, and Bacillus subtilis is typically used in the fermentation of soybean-based foods. The fermentation process may improve not only the flavor but also the nutritional value of food, and substances produced in this fermented broth were affected by many factors including culture medium and the selected soybeans. In this study, we use 3 potential culture mediums in the fermentation of black soybean and the fermented black soybean broths were used for the examination of amino acid composition, total phenolics content, flavonoids and anthocyanins contents, the antioxidant properties, and cytotoxicity. Our results indicated that the fermented black soybean broth, fermentation III, have the most abundant essential amino acid (79.77 mg/g), phenolics (19.33 mg/g), flavonoids (46.01 mg/g), and anthocyanins (1.06 mg/g). Besides, all of the fermented black soybean broths exhibited the significant antioxidative abilities with 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effect, reducing power and ferrous ion chelating effect. In addition, the fermented black soybean broths demonstrated the cell proliferation-enhancing activity in Detroit 551 cells. The cells were augmented up to the maximum value of 183.6% (compared with control) at 10 mg/mL of the fermentation I. Therefore, the different supplemental culture medium fermented black soybean broths may be used as a functional ingredient in the products of nutritional drinks and health foods. PRACTICAL APPLICATION: The present study illustrated the potential of various supplemental culture medium fermented black soybean broths in the application of functional ingredient for nutritional drinks and health foods.     

不同菌株对发酵大豆后熟期间品质特性的影响

为研究利用细菌和霉菌进行混合发酵的不同效果,开发新的混菌发酵工艺,本研究采用不同接种方式,得到3个单一菌株发酵样品,4个混合菌株发酵的样品,研究不同菌株对发酵大豆品质特性的影响。按照韩国清麴酱的方法,利用Bacillus subtilis菌株发酵制得样品B,按照豆豉的发酵方法利用Aspergillus oryzae和Mucor racemosus菌株制得样品A和M。混合菌株发酵样品中依据两种菌株接种的顺序不同,分别制得AB、BA、MB和BM。结果表明,后熟0~9 d期间,样品B的蛋白酶活性最高(257.32±3.04)U/g;样品M的还原糖在后熟第1 d达到1.17%,高于其他样品。样品A和MB氨基态氮含量较高,样品B中测得铵态氮含量最高。所有样品的pH呈下降趋势,而滴定酸度变化范围则相反,从0.02%上升到0.19%。在后熟9 d期间,各样品色度中的L*值呈持续下降趋势,样品MB褐变现象明显。在7个实验组中,样品MB通过混菌发酵工艺提高了发酵大豆的品质特性,验证了利用细菌和霉菌进行混菌发酵技术能够改善发酵大豆的品质特性。     

产谷氨酸脱羧酶重组枯草芽孢杆菌的构建与食品级表达

谷氨酸脱羧酶(GAD)是生物合成γ-氨基丁酸(GABA)的关键酶。本研究构建了4株无抗标记的重组枯草芽孢杆菌,首次实现了乳酸乳球菌(Lactococcus lactis ssp.lactis IL 1403)来源的谷氨酸脱羧酶基因在枯草芽孢杆菌中的食品级表达。通过比较4株重组枯草芽孢杆菌的生长曲线和发酵酶活曲线,筛选出产酶效率最高的重组菌株B.subtilis WB600/pUB-P43-gadB(opt)-dal,该重组菌在初始发酵培养基中发酵42 h后发酵酶活可达4.1 U/mL。通过调整培养基成分,重组菌B.subtilis WB600/pUB-P43-gadB(opt)-dal的发酵酶活最高达到7.4 U/mL,与初始相比酶活提高了79%,是目前已报道重组枯草芽孢杆菌产谷氨酸脱羧酶酶活的最高水平。     

Genotyping of starter cultures of Bacillus subtilis and Bacillus pumilus for fermentation of African locust bean (Parkia biglobosa) to produce Soumbala

Bacillus spp. are the predominant microorganisms in fermented African locust bean called Soumbala in Burkina Faso. Ten strains selected as potential starter cultures were characterised by PCR amplification of the16S-23S rDNA intergenic transcribed spacer (ITS-PCR), restriction fragment length polymorphism of the ITS-PCR (ITS-PCR RFLP), pulsed field gel electrophoresis (PFGE) and sequencing of the 968-1401 region of the 16S rDNA. In previous studies, the isolates were identified by phenotyping as Bacillus subtilis and Bacillus pumilus. The phenotyping was repeated as a reference in the present study.The ITS-PCR and ITS-PCR RLFP allowed a typing at species level. The PFGE was more discriminative and allowed a typing at strain level. Full agreement with the phenotyping was observed in all cases. The sequencing of the 16S rDNA allowed the identification at species level with an identity from 97% to 100% comparing the sequences to those from the GenBank databases. The desired cultures of B. subtilis and B. pumilus from African locust bean fermentation were distinguished by ITS-PCR and ITS-PCR RLFP from Bacillus cereus and Bacillus sphaericus which sometimes occur in the beginning of the fermentation.     

基于酶学特性筛选大曲来源芽孢杆菌用于强化酿酒

通过分析来源于清香型、浓香型和酱香型大曲的105 株芽孢杆菌的产淀粉酶和蛋白酶的能力,优选出1 株地衣芽孢杆菌（Bacillus licheniformis）B.L-1和1 株枯草芽孢杆菌（B. subtilis）B.S-1,将其分别应用于清香型白酒酿造中,探究强化接种微生物对发酵的影响。发酵结束后,在酒醅中共检测到挥发性代谢产物38 种,通过多元统计分析,在强化B.L-1组和B.S-1组中分别筛选出差异代谢物21 个和11 个（VIP＞1;P＜0.05）,其中4-乙基-2-甲氧基苯酚、辛酸乙酯、3-甲基丁酸乙酯和四甲基吡嗪在2 组强化中均为差异代谢物,含量显著升高。结果表明,用酶学特性进行菌株筛选能够快速有效地获得特定功能菌株,由此获得的菌株进行强化发酵可显著提高酒醅中挥发性风味物质含量。该方法可以有效地为大曲微生物合成群落重塑提供菌种资源。