Food Science Commons^™

A Study On Plant-Derived Small Proteins And Peptides With Antiyeast Activity And Preliminary Investigations In Food Materials, Giulia Mignone

Theses

The discovery of novel antimicrobial molecules would benefit considerably different biotechnological fields, including medicine, agriculture and the food industry, on which this thesis work focuses. Specifically, here are presented fundamental studies on plant-derived antimicrobial proteins for the reduction of food spoilage caused by yeast microorganisms. Small proteins and peptides from the plant innate immune system possess several characteristics which could make them ideal candidates for a novel and natural antispoilage agent. The first Chapter reviews proteins families associated with the plant innate immune system that exhibit antiyeast activity and low molecular weight (<30 kDa), focusing on their structure and antiyeast mode of action. Chapter 2 investigates two peptides, Pn-AMP1 and Pn-AMP2 belonging to the hevein-type peptides family; peptides were synthetically generated using their native amino acid sequence. Synthetic Pn-AMP1 showed fast action activity against common food spoilage yeasts, it resulted potentially safe for consumption, and it was successfully incorporated in UHT milk and Fanta Orange, where it contained the growth of Kluyveromyces lactis and Zygosaccharomyces bailii, respectively. A different family of antimicrobial plant proteins (2S albumins) was studied in Chapters 3, 4, 5 and 6. Chapter 3 describes the selective extraction of an antiyeast protein (labelled WMS1 in this thesis work) of ~14 kDa from white mustard (Brassica hirta) seeds. The protein was purified in a relatively high yield, using a simple protocol that included only one chromatographic method step. In Chapter 4, bioinformatics tools and wet-lab techniques are used to study the structure and classify the antiyeast protein. The WMS1 antiyeast protein was identified as an isoform of the Napin protein Allergen Sin a 1 belonging to the 2S albumins family, characterised by high structural stability which donates to the protein resistance heat treatment and proteolytic degradation. Chapter 5 describes the antimicrobial spectrum of WMS1 , its antiyeast mechanism of action and its validity as preservative agent. Although WMS1 resulted potentially safe for consumption and inhibited the growth of Z. bailii in several beverage systems, its biotechnological potential is limited by its allergic properties. In Chapter 6, the protein WMS1 was cloned in a pET28a-MBP vector and expressed in an E. coli BL21 system. Interestedly, recombinant WMS1 and native WMS1 showed similar activity against the yeast Z. bailii. Thesis abstract 2 Chapter 7 includes a protocol for the rational design of ultra-short antimicrobial peptides. Since ultra-short antimicrobial sequences can be found within the sequence of any larger protein, the protocol was applied using WMS1 primary sequence, and seven ultra-short SinA-AMPs were chemically generated. Among the various SinA-AMPs, the peptide labelled SinA-pepIII had in vitro activity against several common yeast spoilage strains via membrane permeabilisation and it could be employed as a food preservative as it was pH- and heat- stable, potentially safe in terms of human consumption and it prevented the growth of Z. bailii in cranberry juice. The results obtained in this work offer new perspectives in the research for novel and clean-label food preservatives to fight the food waste phenomenon.

Exploiting Bacteriophages And Associated Peptidoglycan Hydrolases With Potential For Biocontrol In Food Related Applications, Lorraine Endersen

Theses

The work presented in this thesis describes the isolation and characterisation of novel bacteriophages, and the subsequent exploitation of their bactericidal properties against two opportunistic food related pathogens, Mycobacterium avium subsp. paratuberculosis (MAP) and Cronobacter sakazakii.

MAP is a proven animal pathogen known to cause Johne’s disease in cattle but has been implicated as a causative agent of Crohn’s disease in humans. Despite the fact that this association has yet to be proven, significant focus has been directed towards evaluating the consequences of consuming milk contaminated with MAP. Accordingly, six mycobacteriophages were isolated and characterised in terms of temperature and …

Listeria Monocytogenes'. A Genomic Perspective On Its Pathogenesis, Persistence And Bacteriophages., Aidan Casey

Theses

The objective of this study was to investigate the genetic characteristics of Listeria monocytogenes that contribute to the threat it poses to the food processing industry. Transcriptomic analysis of a persistent L. monocytogenes isolate following exposure to an industrial detergent identified key upregulations in biological processes involved m peptidoglycan biosynthesis, chemotaxis and motility. Two bacteriophages against serotype 4b and 4e isolates of L. monocytogenes were isolated and sequenced. Comparative genomic and functional analysis identified the receptor binding proteins in each phage, whi]e transmission electron microscopy demonstrated a conserved baseplate architecture which also exists in other Gram­positive phages. Comparative genomics of …

Evaluation Of Novel Low Molecular Weight Antimicrobial Compounds Produced By Lactic Acid Bacteria., Jiahui Leona Guo

Theses

Fungal food spoilage plays a pivotal role in the deterioration of food and feed systems and some fungi are also able to produce compounds toxic for humans and animals. The mycotoxins produced by fungi can cause serious health hazards and include toxins which are cancerogenic, immunotoxic, teratogenic, neurotoxic, nephrotoxic or hepatotoxic effects and Kashin-Beck disease. In addition to this, fungal spoilage/pathogens cause losses of marketable quality and hygiene of foodstuffs, resulting in major economic problem throughout the world. Nowadays, food spoilage can be prevented using physical and chemical methods, but no efficient strategy has been proposed so far to reduce …

Development Of Pcr Assays To Detect And Characterise Salmonella Isolates From Cheese And Meat Products By Utilizing An Integrated Genomics Approach, Noelle Geraldine Mccarthy

Theses

Salmonella is an extremely important foodborne pathogen which is responsible for millions of cases of gastroenteritis annually. Salmonella outbreaks have been associated with numerous common food stuffs such as meat, dairy and seafood. Traditional culture based methods for the detection Salmonella are labourious and costly. Rapid detection and serovar identification would dramatically decrease the time required to identify Salmonella outbreaks and potential health risks.

In the US 99% of all Salmonella infections are caused by members of Salmonella enterica subspecies 1. Serovars within subspecies 1 can vary due to host specificity and pathogenicity. DNA sequencing projects have provided valuable genomic …