Department of Food Science and Technology, The Ohio State University, Columbus, Ohio 43210, USA.
J Dairy Sci. 2011 Sep;94(9):4374-82. doi: 10.3168/jds.2011-4457.
The acceptability of Swiss cheese largely depends on the flavor profile, and strain variations of cheese cultures will affect the final quality. Conventional biochemical methods to identify the cultures at the strain level are time-consuming and expensive, and require skilled labor. Our objective was to develop rapid classification methods of starter cultures at the strain level by using a combination of hydrophobic grid membrane filters and Fourier transform infrared (FTIR) spectroscopy. Forty-four pulsed-field gel electrophoresis-verified strains of starter and nonstarter cultures including Streptococcus thermophilus, Propionibacterium freudenreichii, and Lactobacillus spp. were analyzed. The strains were grown on their respective agar media, transferred to broth media, and incubated. Then, cultures were centrifuged and the pellets were resuspended in saline solution (10 μL). Aliquots (2 μL) of the suspended bacterial solution were placed onto a grid of the hydrophobic grid membrane filters, having 6 grids per each strain analyzed. The dried filters were read by FTIR microspectroscopy fitted with an attenuated total reflectance probe. Collected spectra were statistically analyzed by a soft independent modeling of class analogy (SIMCA) for pattern recognition. Classification models were developed for Streptococcus thermophilus, Propionibacterium freudenreichii, and Lactobacillus spp. strains. The models showed major discrimination in the spectral region from 1,200 to 900 cm(-1) associated with signals from phosphate-containing compounds and various polysaccharides in the cell wall. The developed method allowed for rapid classification of several Swiss cheese starter and nonstarter cultures at the strain level. This information provides a detailed overview of microbiological status, which would enable corrective measures to be taken early in the cheese making process, limiting production of inferior quality cheese and minimizing defects. This method could be an effective tool to identify and monitor activity of cheese and other dairy starter cultures.
瑞士干酪的可接受性在很大程度上取决于其风味特征,而奶酪培养物的菌株变化将影响最终质量。传统的生化方法在菌株水平上鉴定培养物既费时又昂贵,并且需要熟练的劳动力。我们的目标是开发一种快速的分类方法,在菌株水平上通过使用疏水性网格膜过滤器和傅里叶变换红外(FTIR)光谱相结合的方法来鉴定起始培养物。分析了 44 株经脉冲场凝胶电泳验证的起始和非起始培养物的菌株,包括嗜热链球菌、丙酸杆菌和乳杆菌属。将这些菌株在各自的琼脂培养基上生长,转移到肉汤培养基中进行培养。然后,将培养物离心,将沉淀重悬于生理盐水(10 μL)中。将 2 μL 的悬浮细菌溶液等分试样放置在疏水网格膜过滤器的网格上,每个分析的菌株有 6 个网格。将干燥的过滤器用配备衰减全反射探针的 FTIR 微光谱仪读取。通过软独立建模分类分析(SIMCA)对收集的光谱进行统计分析,以进行模式识别。为嗜热链球菌、丙酸杆菌和乳杆菌属的菌株开发了分类模型。这些模型在与含磷酸盐化合物和细胞壁中各种多糖相关的 1200 到 900 cm(-1) 的光谱区域显示出主要的区分。所开发的方法允许在菌株水平上快速分类几种瑞士干酪的起始和非起始培养物。该信息提供了微生物状况的详细概述,这将使人们能够在奶酪制作过程的早期采取纠正措施,限制劣质奶酪的生产并最大限度地减少缺陷。该方法可以成为识别和监测奶酪和其他乳制品起始培养物活性的有效工具。