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Lactobacillus delbrueckii ssp. bulgaricus Milk- and Low-Temperature-Associated Proteomes

ABSTRACT: We identified the proteins synthesized by Lactobacillus delbrueckii ssp. bulgaricus strain LBB.B5 in laboratory culture medium (MRS) at 37 �C and in milk at 37 and 4 �C. Cell-associated proteins were measured by gel-free, shotgun proteomics using high-performance liquid chromatography coupled with tandem mass spectrophotometry. A total of 635 proteins were recovered from all cultures, and among which 72 proteins were unique to or significantly more abundant in L. bulgaricus after growth in milk compared to MRS at 37 �C.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Lactobacillus Delbrueckii (ncbitaxon:1584)

SUBMITTER: Maria Marco

PROVIDER: MSV000080650 | MassIVE | Fri Mar 17 17:20:00 GMT 2017

SECONDARY ACCESSION(S): PXD006110

REPOSITORIES: MassIVE

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Project description:Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, the transcriptome derived from the milk fat layer has not been compared with the mammary-derived transcriptome nor have sources of RNA been quantified in milk. In this study the effects of milk collection and processing on RNA quality and origin were assessed in humans and rhesus macaques. Total RNA in milk was quantitated in acridine orange-stained milk using an automated whole slide scanner and custom-built Globulator software. Total RNA extracted from milk fat, cells in milk, and mammary biopsies of lactating rhesus macaques were compared using RNA sequencing and analysis. Compared with human milk, milk from macaques contained similar amounts of RNA-containing cytoplasmic crescents, but more cells. Total RNA extracted from milk fractions was also evaluated for factors that affect RNA quality. Degradation of RNA extracted from human milk fat was positively correlated with geographic distance from collection site, storage time, and sample type. There were no differences in RNA degradation in macaque milk collected after 10 min or 4 hr accumulation, suggesting that degradation of RNA extracted from milk fat may not occur in the mammary gland. Using RNA-Seq, RNA extracted from macaque milk fat and cells in milk more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from mammary epithelial cells. Analysis of highly abundant putative microRNAs in macaque milk fat revealed a potentially novel non-coding RNA species that is conserved in humans. RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells. However, this sample type clearly requires protocols that minimize RNA degradation.

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Lactobacillus delbrueckii ssp. bulgaricus Milk- and Low-Temperature-Associated Proteomes

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