Study of Cheese Associated Lactic Acid Bacteria Under Carbohydrate-Limited Conditions Using D-Stat Cultivation

The metabolic pathways alternative to glycolytic energy (ATP) during growth of starter and nonstarter lactic acid bacteria were studied simulating the depletion of carbohydrates during cheese ripening. D-stat cultivation strategy with the gradual decrease of galactose concentration in tryptone-arginine feeding medium was used. With the decrease of galactose feeding, the biomass yield calculated on carbohydrate consumption (Y_X/HEX) and acetate/lactate production ratio of all strains increased. We assume that ATP and biomass yields improved by directing the pyruvate flow from lactate to acetate and that metabolic energy could be obtained either by producing acetate from carbohydrates or from arginine metabolism in ADI-positive strains. Four LAB strains out of eight produced ornithine from arginine indicating active arginine-deiminase (ADI) pathway. These ADI-positive strains achieved 3-10 times higher Y_X/HEX than ADI-negative strains in tryptone-arginine medium. Lactobacillus plantarum also used serine as an energy source. Starters and NSLAB strains using the amino acids arginine and serine or limited amounts of carbohydrates therefore have the potential to influence flavor production in cheese more efficiently.     

Study of Cheese Associated Lactic Acid Bacteria Under Carbohydrate-Limited Conditions Using D-Stat Cultivation

The metabolic pathways alternative to glycolytic energy (ATP) during growth of starter and nonstarter lactic acid bacteria were studied simulating the depletion of carbohydrates during cheese ripening. D-stat cultivation strategy with the gradual decrease of galactose concentration in tryptone-arginine feeding medium was used. With the decrease of galactose feeding, the biomass yield calculated on carbohydrate consumption (Y_X/HEX) and acetate/lactate production ratio of all strains increased. We assume that ATP and biomass yields improved by directing the pyruvate flow from lactate to acetate and that metabolic energy could be obtained either by producing acetate from carbohydrates or from arginine metabolism in ADI-positive strains. Four LAB strains out of eight produced ornithine from arginine indicating active arginine-deiminase (ADI) pathway. These ADI-positive strains achieved 3–10 times higher Y_X/HEX than ADI-negative strains in tryptone-arginine medium. Lactobacillus plantarum also used serine as an energy source. Starters and NSLAB strains using the amino acids arginine and serine or limited amounts of carbohydrates therefore have the potential to influence flavor production in cheese more efficiently.     

Technological and probiotic role of adjunct cultures of non-starter lactobacilli in soft cheeses

The influence of two cheese-isolated Lactobacillus strains on cheese composition, acceptability and probiotic capacity was assessed. Soft cheeses with and without the addition of Lactobacillus plantarum I91 or Lactobacillus paracasei I90 were prepared. Gross composition was assessed and secondary proteolysis was described by soluble fractions and free amino acids profiles. Acceptability was determined by a panel of 98 non-trained consumers. Cheeses harboring added Lactobacillus strains were also studied in vivo to evaluate their probiotic capacity. Gross composition of the cheeses was similar for control and treated (Lactobacillus-added) cheeses. Peptidolysis increased in cheeses with added lactobacilli, which was evidenced by a higher free amino acid content. Overall, the acceptability of the cheeses was good: 65%–80% of the consumers said that they “liked very much” or “liked” the cheeses. Cheeses with L. plantarum I91 showed the highest changes in composition and proteolysis and were the most accepted ones. On the contrary, composition of cheeses with L. paracasei I90 was similar to that of the controls, but these samples were less accepted than cheeses without lactobacilli. The oral administration of cheese containing L. plantarum I91 or L. paracasei I90 proved to be safe and able to enhance the number of IgA + cells in the small intestine lamina propria of mice. The use of selected strains of NSLAB exerted a technological and probiotic role: it contributed to the standardization of cheese quality and induced benefic health effects at the gut mucosa in vivo.     

Changes in microbial populations, kinds of lactic acid bacteria and biochemical characteristics of Greek traditional feta cheese during ripening

Three batches of feta cheese manufactured from raw (R) and thermized (TS) milk with yogurt as a starter were studied. Enterobacteriaceae and coliforms underwent a more accelerated decrease in TS cheese. Nonstarter lactic acid bacteria (NSLAB) were counted at higher levels in R than in TS cheese throughout ripening and predominated over the other microbial groups. The composition of NSLAB in the fresh cheese was similar for both cheese types. Proteolysis products (noncasein nitrogen soluble in 12% trichloroacetic acid, nitrogen soluble in 5% phosphotungstic acid /100 total nitrogen) were higher in R than TS cheese. Degradation of αs-casein was in R > TS, while a small reduction of β-casein during storage was recorded only for TS cheese.     

干酪促熟附属发酵剂的研究进展

干酪成熟时间较长且费用较高,干酪促熟成为降低生产成本的有效途径之一。干酪促熟常用的方法有酶法、修饰发酵剂细胞、提高成熟温度、高压处理等,但均存在一定的不足,限制了其在干酪工业中的应用。非发酵剂乳酸菌可促进干酪风味的形成并加速成熟．已成为干酪促熟方法的研究热点之一。介绍了干酪生产及成熟过程中微生物的作用,特别介绍了干酪附属发酵剂发展的原由并综述了其在干酪成熟过程中研究进展。     

新疆酸驼乳NSLAB的筛选及鉴定

以新疆牧民家庭自然发酵酸驼乳为原料,采用改良的ROGOSA培养基筛选出20株非发酵剂乳酸菌（Non Starter Lactic Acid Bacteria,NSLAB）,以产酸、自溶度、氨肽酶活力、蛋白水解能力为指标复筛性能优良的NSLAB,为新疆特色干酪的现代化改造提供新的菌株资源。结果表明:从酸驼乳中复筛得到六株NSLAB（编号X-4、X-5、X-7、X-12、X-13、X-15）。据多元统计分析六株菌综合得分X-5>X-4>X-15>X-13>X-12>X-7。其中X-5Δp H（24h）、自溶度、氨肽酶活力、蛋白水解能力分别为1.08%、28.26%、18.88、3.21U/m L,有较好的应用前景。基于菌株生理生化特性,结合16S r DNA基因序列分析,构建菌株系统发育树明确其分类地位,X-4、X-5、X-13、X-15鉴定为植物乳杆菌（Lactobacillus plantarum）,X-7鉴定为发酵乳杆菌（Lactobacillus fermentum）,X-12确定为乳酸片球菌（Pediococcus acidilactici）。      

Cheddar干酪附属发酵剂筛选及其应用

为了筛选能够促进Cheddar干酪成熟,从泡菜、腐乳及不同产地的Cheddar中筛选具有高肽酶活力和自溶度的乳杆菌作为Ched-dar干酪附属发酵剂,并应用于Cheddar干酪的制作中.结果表明,从美国Cheddar干酪中筛选出1株Lactobacillus plantarum OD具有较高的肽酶活力和自溶度,运用于Cheddar千酪的制作中能显著提高其游离氨基酸浓度,改善其风味,有利于加快干酪的成熟.     

Extension of Tosèla cheese shelf-life using non-starter lactic acid bacteria

Six strains of non-starter lactic acid bacteria (NSLAB) were used to extend the shelf-life of the fresh cheese Tosèla manufactured with pasteurised cows’ milk. The acidification kinetics of three Lactobacillus paracasei, one Lactobacillus rhamnosus and two Streptococcus macedonicus were studied in synthetic milk medium. Lb. paracasei NdP78 and NdP88 and S. macedonicus NdP1 and PB14-1 showed an interesting acidifying capacity and were further characterised for growth in UHT milk and production of antimicrobial compounds. Lb. paracasei NdP78 and S. macedonicus NdP1 grew more than 2 log cycles in 6 h. Lb. paracasei NdP78 was also found to produce a bacteriocin-like inhibitory substance (BLIS) active against Listeria monocytogenes. The four NSLAB strains (singly or in combination) were used to produce experimental pilot-scale cheeses which were compared by a panel. The cheese manufactured with the mixed culture Lb. paracasei NdP78 - S. macedonicus NdP1 was the most appreciated for its sensory properties. The cheeses produced at factory-scale showed higher concentrations of lactobacilli (7.90 log CFU/g) and streptococci (6.10 log CFU/g), but a lower development of coliforms (3.10 log CFU/g) and staphylococci (2.78 log CFU/g) than control cheese (4.86, 4.89, 4.93 and 5.00 log CFU/g of lactobacilli, streptococci, coliforms and staphylococci, respectively) processed without NSLAB addition. The food pathogens Salmonella spp. and Listeria monocytogenes were never detected. The dominance of the species inoculated was demonstrated by denaturing gradient gel electrophoresis (DGGE), whereas strain recognition was evaluated by randomly amplified polymorphic DNA (RAPD)-PCR. From the results obtained, Lb. paracasei NdP78 and S. macedonicus NdP1 were able to persist during the storage of Tosèla cheese and their combination influenced positively the sensory characteristics and shelf-life of the final product.