Development of a defined minimal culture medium for Prevotella copri DSM 18205
(2020) KBTM01 20201Biotechnology (MSc)
Biotechnology (M.Sc.Eng.)
- Abstract
- The human gut microbiota is involved in the host’s health and disease. There has been growing interest in the next generation probiotic candidate Prevotella copri, which plays a beneficial role in glucose metabolism. It is a Gram-negative obligate anaerobe that was first isolated from human faeces and which is present in plant-based fermented foods as well as in human breast milk. The aim of this project was to develop a minimal culture medium free from animal-derived ingredients, within the framework of potentially developing a fermented food product that contains P. copri in collaboration with Aventure AB.
The first phase of the project consisted of assessing the optimal pH condition as well as the efficacy of commercial horse serum... (More) - The human gut microbiota is involved in the host’s health and disease. There has been growing interest in the next generation probiotic candidate Prevotella copri, which plays a beneficial role in glucose metabolism. It is a Gram-negative obligate anaerobe that was first isolated from human faeces and which is present in plant-based fermented foods as well as in human breast milk. The aim of this project was to develop a minimal culture medium free from animal-derived ingredients, within the framework of potentially developing a fermented food product that contains P. copri in collaboration with Aventure AB.
The first phase of the project consisted of assessing the optimal pH condition as well as the efficacy of commercial horse serum in the complex medium PYG (Peptone Yeast Glucose) for growing P. copri. The second phase of the project involved the formulation of the DMMG (Defined Minimal Medium Glucose) as well as the evaluation of plant-derived haemoglobin (BvHb) as a substitute of commercial hemin in the culture medium.
The optimal pH for growing P. copri was 7.2, and the addition of horse serum was not indispensable for its successful growth in PYG. The DMMG was developed based on culture media previously formulated for Bacteroides fragilis, which rendered an acceptable biomass and succinic acid production. Moreover, the omission of hemin favoured the growth of P. copri in DMMG, which is inconsistent with previous studies. This finding suggests that more research is needed in order to elucidate the role of hemin in the cultivation of P. copri as well as to improve the formulation of the DMMG to enhance the biomass production. (Less) - Popular Abstract
- Humans have been consuming probiotics without realising it during centuries through traditional fermented foods such as yoghurt and kefir. Later on we have found out that some of these bacteria can be beneficial for the prevention and treatment of conditions such as inflammatory bowel disease or allergies. However, what if probiotics could be used to target or even cure diseases? This is what the so-called Next Generation Probiotics (NGP) are aiming to do, and Prevotella copri is a candidate to become one. P. copri is a bacterium naturally present in the human gut, breast milk, as well as in sauerkraut and beer. But why is P. copri so interesting? Because this bacterium plays a beneficial role in glucose metabolism, since it is present in... (More)
- Humans have been consuming probiotics without realising it during centuries through traditional fermented foods such as yoghurt and kefir. Later on we have found out that some of these bacteria can be beneficial for the prevention and treatment of conditions such as inflammatory bowel disease or allergies. However, what if probiotics could be used to target or even cure diseases? This is what the so-called Next Generation Probiotics (NGP) are aiming to do, and Prevotella copri is a candidate to become one. P. copri is a bacterium naturally present in the human gut, breast milk, as well as in sauerkraut and beer. But why is P. copri so interesting? Because this bacterium plays a beneficial role in glucose metabolism, since it is present in higher numbers in the gut of people who have an improved glucose tolerance after consuming fiber-rich foods. Thus, P. copri has a potential to target metabolic diseases such as type 2 diabetes or obesity. But before this can be a reality, we need to find a way to grow P. copri efficiently.
This project was carried out at the division of Biotechnology of Lund University in collaboration with Aventure AB. The final scope is to develop a vegan fermented product that contains P. copri, and this master thesis was focused on finding a suitable culture medium as a first step. P. copri is usually grown in complex culture media (such as PYG – Peptone Yeast Glucose), which means that a lot of ingredients are needed, and the exact chemical composition of the broth is unknown. Moreover, if the final product is going to be plant-based, the culture medium should also be free from animal-derived ingredients, which is unusual when growing this type of bacteria.
During this project we developed a defined minimal medium, in other words, a simple culture broth for P. copri, free from animal-derived ingredients. The experiments were carried out in small serum bottles and both the growth and the production of bacterial metabolites were monitored. The most important product by P. copri is succinate, which has a beneficial role in glucose metabolism. Different versions of the defined medium were developed, with and without plant-derived haemoglobin. Previous research has shown that hemin (a component found in haemoglobin) is required to grow P. copri, especially in defined medium. However, P. copri reached higher numbers and produced more succinate in the defined medium without haemoglobin, which was unexpected but good news for the aim of the project. Nevertheless, more work is needed in order to enhance the growth of P. copri in defined medium.
To sum up, this project shows that it is possible to grow P. copri in a minimal medium with no animal-derived ingredients, even when omitting hemin. As this hadn’t been achieved before, it is a promising step for developing a vegan fermented product with P. copri in its formulation. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9032474
- author
- Nieto Esteve, Anna LU
- supervisor
- organization
- course
- KBTM01 20201
- year
- 2020
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Prevotella copri, probiotics, fermentation, hemin, biotechnology, bioteknik
- language
- English
- id
- 9032474
- date added to LUP
- 2020-12-11 14:16:56
- date last changed
- 2021-06-30 03:40:04
@misc{9032474,
abstract = {{The human gut microbiota is involved in the host’s health and disease. There has been growing interest in the next generation probiotic candidate Prevotella copri, which plays a beneficial role in glucose metabolism. It is a Gram-negative obligate anaerobe that was first isolated from human faeces and which is present in plant-based fermented foods as well as in human breast milk. The aim of this project was to develop a minimal culture medium free from animal-derived ingredients, within the framework of potentially developing a fermented food product that contains P. copri in collaboration with Aventure AB.
The first phase of the project consisted of assessing the optimal pH condition as well as the efficacy of commercial horse serum in the complex medium PYG (Peptone Yeast Glucose) for growing P. copri. The second phase of the project involved the formulation of the DMMG (Defined Minimal Medium Glucose) as well as the evaluation of plant-derived haemoglobin (BvHb) as a substitute of commercial hemin in the culture medium.
The optimal pH for growing P. copri was 7.2, and the addition of horse serum was not indispensable for its successful growth in PYG. The DMMG was developed based on culture media previously formulated for Bacteroides fragilis, which rendered an acceptable biomass and succinic acid production. Moreover, the omission of hemin favoured the growth of P. copri in DMMG, which is inconsistent with previous studies. This finding suggests that more research is needed in order to elucidate the role of hemin in the cultivation of P. copri as well as to improve the formulation of the DMMG to enhance the biomass production.}},
author = {{Nieto Esteve, Anna}},
language = {{eng}},
note = {{Student Paper}},
title = {{Development of a defined minimal culture medium for Prevotella copri DSM 18205}},
year = {{2020}},
}