Dr. Betenbaugh REM Projects (JHU)

As carbon dioxide (CO2) emissions increasingly rise and global warming is intensifying, the need for an alternative source of energy is in demand. Methylomicrobium alcaliphilum 20ZR is a methanotroph that has the ability to utilize methane and methanol as their main carbon source. By utilizing the catalytic conversion to convert CO2 into methanol and genetically modifying the bacteria to produce value-added chemicals, methanotrophs could pose to be useful in the future. As catalytic conversion utilizes potassium-based buffer as a main component in solution, we adapted M. alcaliphilum 20ZR to grow in potassium-rich media. To measure the overall growth of the bacteria, methanol consumption was measured using NMR and cell concentration was taken using the optical density. The bacteria successfully grew in a media that contained 80% KCl. These results suggest that this bacteria has the potential to grow in high concentrations of KCl and utilize methanol as its main carbon source.

As CO 2 emissions continue to rise and fossil fuel dependency increases the need for healthy biofuel use. Alginate beads are becoming useful due to their ability to encapsulate things for long periods of time. Alginate beads are being used commercially in agriculture cosmetics and even in food. The E.coli was cultured before being encapsulated. Once encapsulated E.coli will measure how much sucrose the E.coli metabolizes. The E.coli was contaminated in LB media so it was recultured in M9 media where glucose concentration levels went down, meaning E.coli can be encapsulated in alginate beads.

Water quality and pollution justify a need for more methods of bioremediation to be engineered. Co-cultures have shown increased levels of growth and nutrient recovery between two organisms in past case studies. In this experiment, microalgae and cyanobacteria were chosen due to similar biological properties that may benefit one another and not compete for the same resources. To test this the optical density (OD) and live microscopy pictures were taken to observe the growth of the cultures. The experiment showed co-cultures to have increased growth in a 14 day period as opposed to monocultures of cyanobacteria. The optical density increased at a greater rate in the co-cultures compared to the monocultures. Therefore, co-cultures are ideal as an efficient and cost-effective process for bioremediation of wastewater.

The world is in need of new, renewable energy sources, and algae may be the solution. Hydrogen gas is an extremely energetic molecule which makes it an excellent fuel source, but conventional means of hydrogen production are costly, energy intensive, and produce high levels of CO2. The goal of this experiment is to design and create a working fuel cell that produces hydrogen gas using the biological processes of Chlorella protothecoides, Chlorella kessleri, Chlamydomonas reinhardtii, and Scenedesmus obliquus, the oxidation-reduction (REDOX) reaction between carbon and platinum, and the hydrolysis of water. The amount of electrons used by each algae sample was measured by comparing the current produced from the fuel cell with different applied potentials. Though it seems C. protothecoides is unable to take in electrons indicating it is not reducing protons to hydrogen gas, the fuel cell still has the potential to work with hydrogen-producing algae species and improved equipment.

Both natural and synthetic zeolite have been used to capture different chemical compounds, but they have yet to be seen as both a lucrative and environmentally friendly replacements for traditional processes. We investigated the use of synthetic zeolite (ZSM-5), and a natural, store-bought zeolite as adsorbents for ammonia. Along with comparing the changes in ammonia concentration between natural and synthetic zeolite, it was still necessary to find the trend of the concentration over time. The efficiency of zeolite was tested in both an untreated wastewater solution provided by Baltimore Polytechnic Institute and a stock ammonia solution. From the experimental data we found that the bacteria in the wastewater may also produce ammonia which interfered with our goal of reducing the concentration of ammonia using zeolite.

As the population of the world increases, the demand for more protein and other nutrients in food also increases. Aquaculture could be a viable way of meeting this need for protein but the current practice of feeding farm raised fish feed derived from wild caught fish is unsustainable. The livability of aquaculture can meet the necessity for protein by sustaining far more efficient resources that will aid the possibility of remarkable foods to organisms. We grew Chlorella vulgaris with additional amino acids to increase protein levels. A model was used to predict which amino acids could be used to escalate the protein content in algae. We chose four amino acids for the model. However, taurine wasn’t in the model’s pathway, which meant that we had to find a way to suit it with other linking amino acids, making every amino acid connect and produce a great an efficient amount of protein. The algae were grown in media with the added substrates, which are the different amino acids that we took to place into the strain of Chlorella vulgaris, for eight days while monitoring cell growth, and the biomass was measured at the end of the eight days. Data indicates that cysteine produced the highest biomass and the highest amount of protein production; however, due to precipitation, it was a false growth. This false growth specifies that cysteine have been contaminated while in the process of it growing with Chlorella vulgaris. Hence, the precipitation of cysteine caused a high reading in protein content and growth rate. We recognized that we needed to break it down to acquire accurate results by using Tris Buffer (a buffer used to maintain pH levels), but it did not work quite well. In contrast, serine and methionine were decent sources of increasing protein content. With the production of this study, we can move forward and measure taurine’s content within algae, which may need high-performance liquid chromatography (HPLC). We also can look forward to what dissolves cysteine, since it is such a difficult substance to break down so that it can fit into our model. The funding for this research was provided by the NSF through the EFRI-REM program under award #1332344.

Cyanobacteria is a group of microorganisms which, when grown in the right conditions, have the capability to cover the entire world's oceans. When they are presented with a large amount of nutrients, they can cause algal blooms which can be detrimental to human health. Algal blooms can be toxic to humans and even cause neurological damage. However there are advantages that can be taken from cyanobacteria that could be used to promote human health, the food industry, cosmetics, biofuel, and many other industries. Exopolysaccharides (EPS), and Released polysaccharides (RPS), are naturally made bundles of polysaccharide sugar chains that are produced by microorganisms. These chains either come attached to one sugar or attached to multiple (homopolysaccharides and heteropolysaccharides),and they have been many applications across commercial and industrial fields. Two strains of Nostoc commune, 7413 and 8109, were tested to increase their polysaccharide production and to determine the possible applications of the polysaccharides they produced. The strains were grown in different environments to increase their polysaccharide production. One strain, 7413, was tested in strong and weak light intensity as well as in a salt stressed environment, while 8109 was only tested in a salt stressed environment. 8109 was only grown in the salt stressed environment due to it’s very delicate environmental needs. Biomass samples were collected on an alternating day schedule, dependent on strain, and measured for biomass and polysaccharide production. These samples proved that the cyanobacteria did have an increase in polysaccharide production for both strains. The salt stress also caused the bacteria to not grow as much,as we expected due to the added stress. Future research will include polysaccharides collection and measurement in a gas chromatography mass spectrometry machine (GCMS) for monosaccharide make up . This will allow us to see the individual monosaccharide sugars that make up the polysaccharide, and help, to determine its industrial use.

Industrial, agricultural and domestic pollutants scar the environment as population and urbanization trends increase, affecting fauna and flora due to the burning of fossil fuels that "produces about 2/3 of sulfur dioxide present in the air" (1. Shah). The objective of this study is to utilize a possible solution, algae since it has the ability to remove toxic pollutants from waterways during growth without creating a secondary pollutant. To measure the growth of algae in poor conditioned wastewater, optical densities will be taken with a spectrophotometer measuring the amount of light that is absorbed by the samples and light that isn't. Taking optical densities, we found that each algal strain showed signs of growth: Scenedesmus acutus f. alternans (SCFA) had the highest growth rate and Chlamydomonas reinhardtii (CC503) had the slowest. This shows that every strain doesn't have similar growth patterns in these conditions in the aquaculture wastewater. Despite the difference of growth rates, the strains Scenedesmus acutus f. alternans (SCFA) and Chlorella sorokiniana (CS-02) were used for Fourier transform infrared readings. The readings showed the fingerprint of the strains which were included in each sample. The Fornier transform infrared readings allowed us to find the lipid productions of each strain in each sample. Since these algal strains produce lipids, the horizon that shows the countless ways algae can be used is expanding. They could even become a biofuel which would decrease the amount of harmful fossil fuel emissions since their lipid products can be converted to biodiesel.

Microalgae has a potential of replacing actual fish as an Omega-3 source for farmed fish. However, commercial production of algae is very limited due to the high cost in carbon sources for algae. The aim of this study is to investigate the effect of different carbon sources on the lipid content of Nannochloropsis sp. to better and quicker produce algae. Ethanol and methanol, cheap carbon sources, were found in other experiments to increase the carbon dioxide intake and to be a possible source of carbon for algae. Much research on Chlorella sp. has been conducted and found that it can accumulate lots of lipids. Different Chlorella sp. were initially cultured to test whether the amount of lipid accumulated with the addition of methanol and ethanol will increase.

Due to the ability to accumulate biomass rapidly through photosynthesis, microalgae have become very useful for production of nutritional supplements like lutein, which can help prevent and slow the progression of cataracts. Microalgal oils can also be extracted to make biofuels for cars and more pertinent to the immediate goal of this project, microalgae are a sustainable food source for fish grown at commercial scale. The primary objectives of this study are to determine which level of carbon dioxide is optimal for algal growth, and the amount of lutein and total cell protein yielded by Chlorella vulgaris when cultured with different CO₂ levels. The different CO₂ conditions in this experiment are 0.4% (atmospheric), 5%, and 10%. For the trials testing 0.4% and 5% CO₂ conditions, we detected significantly greater biomass and growth rate with 5% CO₂. We are monitoring the amount of lutein and protein yielded under each condition and these will be reported. Eventually our results will be used to determine which of the three CO2 conditions will be used to grow C. vulgaris as fish food for Tilapia sp. at Baltimore Polytechnic Institute’s aquaponics system.

About 1 billion people rely on fish as their primary source of protein. Another million people consume fish at least once a week. Fish is an essential part of many human diets. Most species of fish have large numbers of polyunsaturated fatty acids which supplement many humans with healthy fats such as, omega 6. These high levels of omega 6 levels are a blessing and a curse. Many modern day farm raised fish are heavy in omega 6 but lack in omega 3. These unbalanced ratios of omega 6 and omega 3 are very unhealthy for the human heart. Due to the fact that many humans consume fish many researchers are searching for a solution for this problem. One promising solution is to create a very nutritious fish food. A promising fish food is algae. My research aimed to discover which algae growth media fostered the highest Omega 3 yield in Nannochloropsis sp. Nannochloropsis sp. has been proven to accumulate high levels of Eicosapentaenoic acid (EPA). Therefore, we decided to grow Nannochloropsis sp. in three different growth media; BG11, BBM, F/2; in order to discover which media would foster the highest EPA yields. The results suggest that Nannochloropsis sp. fosters the highest EPA yields when grown in the F/2 media. We plan to use these results in order to create an algae based fish food; which in turn would increase the nutritional value of many species of fish. Therefore, balancing the ratio of omega 6 and omega 3 levels in fish; which in turn could possibly increase the health of the heart of the billions of people who have fish in their diets.