Guarcello Rosa, De Angelis Maria, Settanni Luca, Formiglio Sabino, Gaglio Raimondo, Minervini Fabio, Moschetti Giancarlo, Gobbetti Marco
Dipartimento di Scienze Agrarie e Forestali, Università degli Studi di Palermo, Palermo, Italy.
Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi di Bari Aldo Moro, Bari, Italy.
Appl Environ Microbiol. 2016 Nov 9;82(23):6870-6880. doi: 10.1128/AEM.01051-16. Print 2016 Dec 1.
Accumulation of biogenic amines (BAs) in cheese and other foods is a matter of public health concern. The aim of this study was to identify the enzyme activities responsible for BA degradation in lactic acid bacteria which were previously isolated from traditional Sicilian and Apulian cheeses. The selected strains would control the concentration of BAs during cheese manufacture. First, 431 isolates not showing genes encoding the decarboxylases responsible for BA formation were selected using PCR-based methods. Ninety-four out of the 431 isolates degraded BAs (2-phenylethylamine, cadaverine, histamine, putrescine, spermine, spermidine, tyramine, or tryptamine) during cultivation on chemically defined medium. As shown by random amplification of polymorphic DNA-PCR and partial sequencing of the 16S rRNA gene, 78 of the 94 strains were Lactobacillus species (Lactobacillus casei, Lb. fermentum, Lb. parabuchneri, Lb. paracasei, Lb. paraplantarum, and Lb. rhamnosus), Leuconostoc species (Leuconostoc lactis and Ln. mesenteroides), Pediococcus pentosaceus, Lactococcus lactis, Streptococcus species (Streptococcus gallolyticus and S. thermophilus), Enterococcus lactis, and Weissella paramesenteroides A multicopper oxidase-hydrolyzing BA was purified from the most active strain, Lb. paracasei subsp. paracasei CB9CT. The gene encoding the multicopper oxidase was sequenced and was also detected in other amine-degrading strains of Lb. fermentum, Lb. paraplantarum, and P. pentosaceus Lb. paracasei subsp. paracasei CB9CT and another strain (CACIO6CT) of the same species that was able to degrade all the BAs were singly used as adjunct starters for decreasing the concentration of histamine and tyramine in industrial Caciocavallo cheese. The results of this study disclose a feasible strategy for increasing the safety of traditional cheeses while maintaining their typical sensorial traits.
Because high concentrations of the potentially toxic biogenic amines may be found in traditional/typical cheeses, the safety of these food items should be improved. Lactic acid bacteria selected for the ability to degrade biogenic amines may be used during cheese making to control the concentrations of biogenic amines.
奶酪及其他食品中生物胺(BAs)的积累是一个备受公众健康关注的问题。本研究的目的是鉴定从西西里和阿普利亚传统奶酪中分离出的乳酸菌中负责生物胺降解的酶活性。所选菌株将在奶酪制作过程中控制生物胺的浓度。首先,使用基于PCR的方法筛选出431株未显示编码负责生物胺形成的脱羧酶基因的分离株。在化学成分确定的培养基上培养时,431株分离株中有94株能降解生物胺(2-苯乙胺、尸胺、组胺、腐胺、精胺、亚精胺、酪胺或色胺)。通过随机扩增多态性DNA-PCR和16S rRNA基因的部分测序表明,94株菌株中有78株为乳杆菌属(干酪乳杆菌、发酵乳杆菌、布氏副乳杆菌、副干酪乳杆菌、类植物乳杆菌和鼠李糖乳杆菌)、明串珠菌属(乳酸明串珠菌和肠膜明串珠菌)、戊糖片球菌、乳酸乳球菌、链球菌属(解脲链球菌和嗜热链球菌)、乳酸肠球菌和副肠膜魏斯氏菌。从活性最高的菌株布氏副干酪亚种副干酪CB9CT中纯化出一种水解生物胺的多铜氧化酶。对编码多铜氧化酶的基因进行了测序,并且在发酵乳杆菌、类植物乳杆菌和戊糖片球菌的其他生物胺降解菌株中也检测到了该基因。布氏副干酪亚种副干酪CB9CT和同一物种的另一株能够降解所有生物胺的菌株(CACIO6CT)被单独用作辅助发酵剂,以降低工业卡乔卡瓦洛奶酪中组胺和酪胺的浓度。本研究结果揭示了一种在保持传统奶酪典型感官特性的同时提高其安全性的可行策略。
由于在传统/典型奶酪中可能发现高浓度的潜在有毒生物胺,因此应提高这些食品的安全性。筛选出具有降解生物胺能力的乳酸菌可在奶酪制作过程中用于控制生物胺的浓度。