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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Exploration Of The Active Site Specificity Of Mala, A Glucosidase From The Predatory Bacterium Bdellovibrio Bacteriovorus, Christine Isabella Jan 2011

Exploration Of The Active Site Specificity Of Mala, A Glucosidase From The Predatory Bacterium Bdellovibrio Bacteriovorus, Christine Isabella

Summer Research

Sequencing of the HD100 genome of Bdellovibrio Bacteriovorus in 2005 revealed a gene for the putative maltase, MalA. However, given the bacterium’s observed disuse of prey carbohydrates as an energy source, this enzyme is seemingly out of place. In this study, the specificity and activity of MalA were explored through various enzymatic assays. Using p-nitrophenol-α-D-glucopyranoside (p-NPG) as a colorimetric substrate to allow for rapid and accurate detection of enzymatic activity through spectrophotometry, enzyme stability inhibition of p-NPG cleavage were explored. While numerous alpha-linked disaccharides were shown to inhibit MalA, only maltose was shown to be cleaved into glucose.


Investigating The Bacterial Predator Bdellovibrio’S Ability To Degrade Aspartate, Scott Anderson Jan 2011

Investigating The Bacterial Predator Bdellovibrio’S Ability To Degrade Aspartate, Scott Anderson

Summer Research

Bdellovibrio bacteriovorus is a predatory Gram-negative Deltaproteobacterium that attacks and invades larger Gram-negative bacteria devouring them from within (Sockett, 2004). Enzymatic results obtained in the 1970s suggest that Bdellovibrio relies on its tricarboxylic acid (TCA) cycle and the oxidation of prey cell derived amino acids (Hespell, 1976). However, annotation of the published genome of Bdellovibrio HD100 revealed that it lacked numerous genes involved with the degradation of amino acids (Rendulic, 2004). Thus it is of great interest to determine if Bdellovibrio can degrade amino acids. If it can, new genes related to the degradation of amino acids will be discovered ...


Developing A Protocol For Purifying The Mala Enzyme In Bdellovibrio Bacteriovorus, John Jared Trecker Jan 2011

Developing A Protocol For Purifying The Mala Enzyme In Bdellovibrio Bacteriovorus, John Jared Trecker

Summer Research

The sequenced genome of the gram-negative predatory bacterium Bdellovibrio bacteriovorus contains a gene that encodes for malA, a putative maltase. Given the bacterium's observed disuse of prey carbohydrates, the gene's presence is mysterious. That characterization of the enzyme and studies of its activity and specificity can be better carried out, it is necessary to obtain pure enzyme. Protein was collected from lysed cultures of Top10/pmalA E. coli. Attempted purification by ion-exchange chromatography with DEAE columns produced significantly purer protein; SP ion exchange columns were unsuccessful, as were heparin and hydroxyapatite affinity columns. Gel filtration chromatography should prove ...


Methods Of Protein Characterization, Michael Tieu Jan 2011

Methods Of Protein Characterization, Michael Tieu

Summer Research

AppA is a protein in Rhodobacter sphaeroides that has been the topic of debate among scientists for the past several years with regards to the structure of the protein. It has been known that AppA has an effect on the activity of PpsR, which controls the gene expression of photosystems. There are two conflicting experimental structures (2IYG and 1YRX) of the protein, both of which claim to be taken in the dark phase (meaning when there is no light shining on the protein). The debate is about whether some slight differences in the structures represent the shift from the dark ...