Molecular interactions of probiotics with oligosaccharides, plant phenolics, and mucin: Subproteome, adhesion, and protein function analyses
Başlık çevirisi mevcut değil.
- Tez No: 403437
- Danışmanlar: Prof. BIRTE SVENSSON, Prof. MAHER ABOU HACHEM
- Tez Türü: Doktora
- Konular: Biyoteknoloji, Mikrobiyoloji, Moleküler Tıp, Biotechnology, Microbiology, Molecular Medicine
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2016
- Dil: İngilizce
- Üniversite: Technical University of Denmark
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 186
Özet
Özet yok.
Özet (Çeviri)
The role of human diet in human health is getting more attention day by day as the environmental changes, especially food contaminations can lead to serious diseases such as cancer. Thus, research is focusing on functional foods such as probiotics and prebiotics in order to promote human health and prevent diseases. Probiotics are microorganisms that exert positive health benefits to the host when administered in sufficient amount and prebiotics are indigestible food ingredients that are selectively fermented by gastrointestinal microbiota, resulting in positive health effects. Furthermore, in recent years, phenolic compounds derived from plant sources have become a very interesting and important subject in human nutrition and health research because they confer health benefits, especially in prevention of cancer and chronic diseases such as cardiovascular diseases and diabetes. This PhD project aims to investigate the interactions between probiotic Lactobacillus acidophilus NCFM (NCFM), approved or emerging prebiotic carbohydrates, plant phenolic compounds, and mucin by i) in vitro analysis of adhesion to mucin and human intestinal cells, ii) comparative wholecell and surface proteomics, and iii) protein functional characterization. In vitro adhesion studies have been used for simulating the in vivo adhesion as adhesion is a crucial property for probiotics to exert their health benefits relating to their residing in the gastrointestinal tract (GIT). Adhesion analysis was performed after growth using nine carbon sources including approved and emerging prebiotics, mucin-supplemented culture, and supplementation with five phenolic compounds. Raffinose, cellobiose, and polydextrose notably increased the adhesion to mucin and to HT-29 cells. Fructooligosaccharides did not change adhesion to mucin, but adhesion to HT-29 cells was increased. Mucin-supplemented growth increased adhesion to HT-29 cells, without changing adhesion to mucin. Adhesion changes by phenolics were dependent on which phenolic was used, as well as their concentration. At 100 μg/mL, resveratrol increased the adhesion to mucin and HT-29 cells; tannic acid increased the adhesion to HT-29 cells; ferulic acid, however, decreased the adhesion to mucin. At 250 μg/mL, resveratrol increased adhesion to HT-29 cells; tannic acid decreased adhesion to mucin; caffeic acid increased the adhesion to mucin; and ferulic acid increased the adhesion to mucin and HT-29 cells. At 500 μg/mL, ferulic acid increased the adhesion to mucin. A 2-D based proteomics strategy was used to identify differentially abundant proteins in response to carbohydrates, mucin, or phenolic compounds. Surface layers (S-layers) are outermost proteinaceous layer of lactobacilli that are responsible for bacteria-host interactions, including adhesion and immune response. Comparison of surface proteomes of NCFM revealed higher abundant surface proteins under different conditions (i.e. growth on raffinose, cellobiose, or mucin-supplemented media) suggesting these proteins are connected with the increased adhesion. Furthermore, whole-cell proteome comparisons of NCFM grown in the presence of plant polyphenols showed that bacteria respond to the polyphenols by altering the metabolism such as the glycolysis and nucleotide metabolism, as well as stress response. Surface proteome comparisons of NCFM grown in the presence and absence of plant polyphenols revealed that the adhesion changes triggered by the polyphenols are correlated with the alterations in relative abundancies of the surface proteome. Four surface proteins were selected for cloning and recombinant production based on differential surface proteome analyses: phosphate starvation inducible protein; pyruvate kinase; elongation factor G (EF-G); and fructose bisphosphate aldolase. Among them, pyruvate kinase and EF-G are previously defined as moonlighting proteins for other bacteria, while the other two are considered as emerging moonlighting proteins for NCFM. Successful cloning and recombinant productions of these proteins were achieved in E. coli; however, over-production of phosphate starvation inducible protein and fructose bisphosphate aldolase has not yet been successful. Pyruvate kinase was over-produced but the yield in purification was not high probably due to the His-tag being inaccessible. Even though the yield was not high, the recombinant pyruvate kinase was used in mucin binding assays and the result showed that the recombinant pyruvate kinase can bind to mucin. In conclusion, the present PhD project adds to the understanding at the molecular level of synbiotic interactions involving NCFM and defined or emerging prebiotics, as well as dietary plant polyphenols. This contributes to understanding of potential positive effects of these combinations on the host originating from alteration in the metabolism and adhesion properties of NCFM.
Benzer Tezler
- Recombinant production of a glycosidase by using an in-vivo cloning system and its immobilization via different strategies
Hücre içi klonlama sistemi kullanarak rekombinant glikosidaz üretimi ve immobilizasyonu
BURCU PEKDEMİR
Yüksek Lisans
İngilizce
2020
BiyokimyaÇanakkale Onsekiz Mart ÜniversitesiBiyomoleküler Bilimler Ana Bilim Dalı
DOÇ. DR. SERCAN KARAV
- Functional chocolate development: Enrichment of dark chocolate with nano-liposome encapsulated antioxidants, and/or pro- and pre-biotics, bioavailability studies
Fonksiyonel çikolata geliştirilmesi: Bitter çikolatanın nano-lipozomla enkapsüle edilen antioksidanlarla, ve/veya pro- ve pre-biyotiklerle zenginleştirilmesi, biyoyararlılık çalışmaları
MİNE ÖZGÜVEN
Doktora
İngilizce
2014
Gıda Mühendisliğiİstanbul Teknik ÜniversitesiGıda Mühendisliği Ana Bilim Dalı
PROF. DR. BERAAT ÖZÇELİK
- Valorization of carrot processing by-products through fermentation and extrusion for enhanced bioactive content and bioaccessibility
Havuç işleme yan ürünlerinin fermentasyon ve ekstrüzyon yoluyla değerlendirilmesi, biyoaktif içerik ve biyoerişilebilirliğin artırılması
DAMLA EZGİ UZUN
Doktora
İngilizce
2025
Gıda Mühendisliğiİstanbul Teknik ÜniversitesiGıda Mühendisliği Ana Bilim Dalı
PROF. DR. ESRA ÇAPANOĞLU GÜVEN
- A computational investigation of the interaction of HCl with small interstellar gas molecules by using quantum mechanical methods
Kuantum mekanik yöntemler kullanılarak HCl molekülünün yıldızlararası ortamdaki küçük gaz molekülleriyle etkileşiminin kuramsal incelenmesi
DUAA ALIA
Yüksek Lisans
İngilizce
2025
Kimyaİstanbul Teknik ÜniversitesiKimya Ana Bilim Dalı
Assist. Prof. BERKAY SÜTAY
- Probiyotik Bifidobacterium breve 'nin bağışıklık düzenleyici tır proteinin tanımlanması, klonlanması ve rekombinant üretimi
Identification, cloning and recombinant expression of immune modulatory tir protein from probiotic Bifidobacterium breve
DİCLE DİLARA AKPINAR
Yüksek Lisans
Türkçe
2019
Gıda MühendisliğiEge ÜniversitesiGıda Mühendisliği Ana Bilim Dalı
DR. ÖĞR. ÜYESİ BURCU KAPLAN TÜRKÖZ