8    MOLECULAR TOOLS FOR
MONITORING BIOREMEDIATION POTENTIAL
ON CONTAMINATED SOILS
Moo Hoon Kim, Senior Researcher
Energy & Environment Lab
Materials & Devices Center
Samsung Advanced Institute of Technology
Suwon, Korea
Lenore S. Clesceri, Associate Professor
Dept. of Biology and Environmental Engineering
Rensselaer Polytechnic Institute
Troy, New York 12180
INTRODUCTION
Cleaning up contaminated soils and sediments is one of the most difficult problems found at
hazardous waste sites. No current technologies are adequate to handle soil cleanup, usually because of cost involved or the unsuitability of the technology to the site environment. Soils at industrial sites are contaminated with wide mixtures of pollutants. Isolation of DNA from contaminated soil has become a very useful tool to study functions in microbial systems. In the next
several years, this procedure will be very important when the DNA probes are available to see
any enzymatic activities. In this research, the bioremediation potential of contaminated soils, the
effectiveness of relationship between lysozyme and SDS treatment and DNA yield by UV
method and recovery from agarose gel electrophoresis were studied and will be presented.
BACKGROUND
Microbial cell wall polysaccharide is made up of two kinds of sugar: N-acetylglucosamine
(NAG) and N-acetylmuramate (NAM). NAM and NAG are derivatives of glucosamine in which
the amino group is acetylated. In bacterial cell walls, NAM and NAG are joined by glucosidic
linkages between C-l of the sugar of NAM and C-4 of the sugar of the NAG. Thus the cell wall
polysaccharide is an altering polymer of NAM and NAG joined by B (1-4) glycosidic linkages.
Lysozyme, a glycosidase, hydrolizes the glycosidic bonds between C-l of the NAM and C-4 of
the NAG. The other glycosidic bond between C-l of NAG and C-4 of the NAM is not cleaved.
Lysozyme is a relatively small enzyme (14.6 kd). The one from egg white, a rich source, is a single polypeptide chain of 129 residues. The gram-positive cell wall has been found to be more
susceptible to lysis with lysozyme than gram-negative wall. Augmenting the lysis mixtures with
detergent (SDS) improves the lysis of gram-positive as well as gram-negative cells.
MATERIALS AND METHODS
Modified direct DNA extraction and purification method (M/H method) from Ogram et al.
(1987) has some disadvantages such as: (1) needs a lot of time, (2) requires a large sample size
so that cannot do the experiment with small sample size, (3) is laborious. Therefore, the rapid
method (E/E method) was tested to overcome larger sample size and save time.
51st Purdue Industrial Waste Conference Proceedings, 1996. Ann Arbor Press. Inc.. Chelsea. Michigan 48118.
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