Identification, typing and characterisation of Propionibacterium strains from healthy mucosa of the human stomach

Forty two Propionibacterium isolates were recovered from biopsy samples of the gastric mucosa of eight out of 12 healthy people. Of these, 41 were identified as belonging to Propionibacterium acnes; the remaining isolate was identified as belonging to Propionibacterium granulosum. Repetitive extragenic palindromic (REP)-PCR typing suggested that up to four strains might be present in the mucosa of the same individual. Sequence analysis of either recA, tly or camp5 genes of P. acnes isolates revealed two distinct phylogenetic lineages. As per the recA, most isolates belonged to type I, while the remainder of the isolates belonged to type II. Phenotypic analyses of representative isolates showed the different strains to have diverse biochemical properties. For example, large differences were seen in carbohydrate fermentation patterns, the results of qualitative and quantitative enzymatic profiling, and survival at acidic pH. In contrast, the patterns of resistance/susceptibility to a series of 16 antibiotics were rather similar, with no atypical resistances observed. The examined strains showed limited-if any-enzymatic activities that could be ultimately related to pathogenicity (lipolytic, proteolytic or haemolytic activity). This suggests that, in the gastric ecosystem, some Propionibacterium spp. genotypes and/or phenotypes can be considered true commensals.     

西南菌种站部分酿酒增香细菌产酸性能初探

本文主要介绍西南菌种站保藏的部分酿酒增香细菌在不同发酵培养条件下产有机酸能力。试验通过对13株菌株进行比较，结果表明，多数丙酸杆菌在合成液培养基中发酵产酸能力较强，丁酸杆菌在母糟浸液培养基中发酵产酸能力较强，菌株SICC1．314、SICC1．259等不仅产酸较高，而且产酸速度也较快。     

Evaluation of in vitro antimicrobial activity of Thai basil oils and their micro-emulsion formulas against Propionibacterium acnes

The aim of this study was to evaluate the efficacy of Thai basil oils and their micro-emulsions, on in vitro activity against Propionibacterium acnes. An agar disc diffusion method was employed for screening antimicrobial activity of the essential oils of Ocimum basilicum L. (sweet basil), Ocimum sanctum L. (holy basil) and Ocimum americanum L. (hoary basil) against P. acnes. Minimum inhibitory concentration (MIC) values of the basil oils were determined using an agar dilution assay. The obtained results indicated that the MIC values of sweet basil and holy basil oils were 2.0% and 3.0% v/v, respectively, whereas hoary basil oil did not show activity against P. acnes at the highest concentration tested (5.0% v/v). Gas chromatography-mass spectrometry analysis revealed that methyl chavicol (93.0%) was the major compound in sweet basil oil, and eugenol (41.5%), gamma-caryophyllene (23.7%) and methyl eugenol (11.8%) were major compounds in holy basil oil. Hoary basil oil contained high amounts of geraniol (32.0%) and neral (27.2%) and small amounts of methyl chavicol (0.8%). The Oil-in-water (o/w) micro-emulsions of individual basil oils with concentrations corresponding to their MIC values were formulated. The stable o/w micro-emulsion system for basil oil consisted of 55.0% v/v water phase, 10.0% v/v oil phase (2.0 or 3.0% v/v sweet basil or 3.0% v/v holy basil oil plus 7.0% v/v isopropyl myristate), 29.2% v/v polysorbate 80 and 5.8% v/v 1,2-propylene glycol. Hydroxyethylcellulose at a concentration of 0.5% w/v was used as thickening agent. According to the disc diffusion assay, the formulations containing sweet basil oil exhibited higher activity against P. acnes than those containing holy basil oil, and the thickened formulations tended to give a lower activity against P. acnes than the non-thickened formulations. The prepared micro-emulsions were stable after being tested by a heat-cool cycling method for five cycles. These findings indicate the possibility to use Thai sweet and holy basil oil in suitable formulations for acne skin care.     

丙酸杆菌的研究进展及其在食品发酵工业中的应用

主要介绍了丙酸杆菌的特性、分类以及其在食品发酵工业中的应用。其应用非常广，主要应用于丙酸、维生素和杆菌素的生产，并简单介绍了它在其他方面的应用.     

维生素B_(12)的生物合成研究

维生素B12广泛应用于医药、食品和畜牧业,主要由微生物发酵得到。脱氮假单胞菌(Pseudomonas denitrificans)和费氏丙酸杆菌(Propionibacterium freudenreichii)是主要的生产菌种。与之相应,维生素B12存在好氧和厌氧2条生物合成途径,生物合成过程十分复杂,2条合成途径大体相同又各有特点。维生素B12发酵产量的提高有待于菌种的改良和发酵工艺的改进。了解维生素B12生物合成途径和代谢调控机制具有重要意义,可为育种工作提供理论基础。     

Capsular exopolysaccharide biosynthesis gene of Propionibacterium freudenreichii subsp. shermanii

In the dairy industry, exopolysaccharides (EPS) contribute to improving the texture and viscosity of cheese and yoghurt and also receive increasing attention because of their beneficial properties for health. For lactic acid bacteria, the production of EPS is well studied. However, for dairy propionibacteria the biosynthesis of EPS is poorly documented. A polysaccharide synthase-encoding gene was identified in the genome of Propionibacterium freudenreichii subsp. shermanii TL 34 (CIP 103027). This gene best aligns with Tts, the polysaccharide synthase gene of Streptococcus pneumoniae type 37 that is responsible for the production of a beta-glucan capsular polysaccharide. PCR amplification showed the presence of an internal fragment of this gene in twelve strains of P. freudenreichii subsp. shermanii with a ropy phenotype in YEL+ medium. The gene sequence is highly conserved, as less than 1% of nucleotides differed among the 10 strains containing the complete gtf gene. The same primers failed to detect the gene in Propionibacterium acidipropionici strain TL 47, which is known to excrete exopolysaccharides in milk. The presence of (1-->3, 1-->2)-beta-d-glucan capsule was demonstrated for 7 out of 12 strains by agglutination with a S. pneumoniae-type 37-specific antiserum. The presence of mRNA corresponding to the gene was detected by RT-PCR in three strains at both exponential and stationary growth phases. This work represents the first identification of a polysaccharide synthase gene of P. freudenreichii, and further studies will be undertaken to elucidate the role of capsular EPS.     

一株费氏丙酸杆菌生长特性及其代谢物抑菌作用分析

对一株费氏丙酸杆菌生长特性及其代谢物抑菌作用进行研究。结果表明：该菌株生长周期约为10 d,第1～3天为其对数生长期,第4～8天为菌体生长的稳定期,第9～10天为衰亡期。在生长过程中,菌体利用糖的速率很快,还原糖在发酵1 d内几乎被消耗殆尽,发酵液pH值在发酵的第1天下降到4.6左右,此后一直到第10天,pH值基本稳定在4.6左右。在同等条件下,该丙酸杆菌代谢物对大肠杆菌的抑制作用优于对金黄色葡萄球菌和酵母菌的抑制作用。对代谢产物的分析结果表明,发酵产物中抑菌物质除了丙酸之外,还存在其他物质,初步判定为细菌素。     

Aerobic culture of Propionibacterium freudenreichii ET-3 can increase production ratio of 1,4-dihydroxy-2-naphthoic acid to menaquinone

This is the first report on the production of both 1,4-dihydroxy-2-naphthoic acid (DHNA) and menaquinone by Propionibacterium freudenreichii ET-3. DHNA can be a stimulator of bifidogenic growth, and menaquinone has important roles in blood coagulation and bone metabolism. During anaerobic culture, DHNA and menaquinone concentrations reached 0.18 mM and 0.12 mM, respectively. The molar ratio between these products was approximately 3:2, which was not affected by culture pH and temperature over the ranges of 6.0-7.0 and 31-35 degrees C, respectively. As for organic acid, propionate and acetate accumulated at concentrations of 0.3 M and 0.15 M, respectively, and the propionate accumulation particularly inhibited further production of DHNA. To improve DHNA production, we switched from anaerobic condition to aerobic condition during the culture when lactose was depleted. DHNA concentration continued to increase even after lactose exhaustion, reaching 0.24 mM. In contrast to DHNA production, menaquinone production stopped after the switch to aerobic condition. The total molar production of DHNA and menaquinone was 0.3 mM irrespective of aerobic culture and anaerobic-aerobic switching culture. Therefore, the anaerobic-aerobic switching culture could increase the production ratio of DHNA to menaquinone. The DHNA concentration obtained from the anaerobic-aerobic switching culture was 1.3-fold higher than that in the anaerobic culture, because P. freudenreichii ET-3 utilized propionate accumulated in the medium via the reversed methylmalonyl CoA pathway under aerobic condition. The culture method proposed in this study could be applicable to industrial-scale fermentation using 1000 l of media, by which 0.23 mM DHNA was produced.