Enzyme-producing strain of bacillus bacteria

Liquid purification or separation – Processes – Treatment by living organism

Reexamination Certificate

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C210S600000, C210S611000, C435S252400, C435S252500, C435S264000, C435S832000, C435S821000, C435S267000, C426S061000, C426S442000, C426S807000, C424S093460

Reexamination Certificate

active

06177012

ABSTRACT:

FIELD OF THE INVENTION
This invention consists of a newly discovered, novel strain of Bacillus bacteria which produce multiple useful enzymes. The novel bacterium and the enzymes it produces have a number of applications, including wastewater treatment, agricultural uses, laundry and dish detergents, drain cleaners and spot removers, among others. In particular, the Bacillus strain of the present invention produces lipase enzymes for the degradation of oleaginous materials such as fats, greases and cooking oils, as well as protease enzymes to degrade proteins and amylases to break down starch.
BACKGROUND OF THE INVENTION
The use of bioaugmentation, i.e., the addition of non-indigenous bacteria to a wastewater to effect more complete treatment and purification of the water, is becoming more and more necessary as municipal sewage plants age and demands brought on by increasing loads require improvements in efficiency. Bioaugmentation, also called bacterial augmentation, can be beneficial in a number of applications. For example, bacterial augmentation can improve the quality and efficiency of treatment of municipal wastewater, food processing wastewater, and residential wastewater, especially in on-site disposal systems such as septic tanks and cesspools, pretreatment of wastewater that might contain high levels of oleaginous matter, and treatment of pipes, traps, plumbing systems and grease traps.
Enzyme-producing bacteria also have agriculture uses as silage inoculant products to improve efficiency and production of livestock that consume the treated silage. Certain bacteria, called Direct Fed Microbials, may also be added to livestock feeds to increase food digestion and utilization. Currently both
Bacillus subtilis
and
Bacillus pumilus
have were found by the Food and Drug Administration to present no safety concerns when used in direct-fed microbial products, as published by the Association of American Feed Control Officials (AAFCO). 1999 Official Publication of the Association of American Feed Control Officials, p. 221. In addition, advances in bioengineering have demonstrated that a genetic characteristic from one species, including bacteria, can be inserted into the DNA of another species. One important example is the insertion of the gene responsible for the production of an insecticidal protein from
Bacillus thuringiensis
into the DNA of corn seeds. The resultant corn plant is able to produce a protein which kills insect pests such as the corn rootworm, thereby producing healthier corn plants and improved per-acre yields.
SUMMARY OF THE INVENTION
The object of this invention therefore is to provide a novel strain of bacteria which produces specific types of useful enzymes.
It is a further object of the present invention to provide a novel strain of bacteria that produces lipase enzymes to degrade fat, proteases to degrade proteins, and amylases to degrade starch.
It is another object of the present invention to provide a novel multiple-enzyme-producing bacterial strain that is useful for wastewater applications to enhance degradation of the organic components of the wastewater and therefore improve the quality and efficiency of the water treatment process.
It is yet a further object of the present invention to provide a novel multiple-enzyme-producing strain of bacteria for use in agricultural applications, including as Direct Fed Microbials in livestock feed formulations, as silage innoculants and to treat livestock manure.
It is a still further object to provide novel bacteria and/or enzymes produced from such bacteria for use as detergents, and to provide enzymes to use as stain removers or in food processing applications.
The present invention provides a novel stain of Bacillus bacteria that produces lipases to degrade fat, proteases to degrade proteins, and amylases to degrade starch. The novel strain disclosed and claimed in the present application is one of eight novel strains invented by the present inventors. For completeness of description, the specification will describe all eight strains, but only the Bacillus strain designated as ATCC 202135 is claimed in the present application. The eight novel strains of Bacillus bacteria have all been deposited with the American Type Culture Collection (“ATCC”), 10801 University Blvd., Manassas, Va., 20110-2209, U.S.A., on Jun. 5, 1998, under the rules of the Budapest Treaty and are designated as follows:
Species
Strain
ATCC Number
Bacillus pumilus
RLM-002
ATCC 202136
Bacillus subtilis
RLM-007Aa
ATCC 202138
Bacillus amyloliquefaciens
RLM-007Ab
ATCC 202134
Bacillus macerans
RLM-007C
ATCC 202132
Bacillus subtilis
RLM-011
ATCC 202137
Bacillus subtilis
RLM-012A
ATCC 202139
Bacillus amyloliquefaciens
RLM-012B
ATCC 202133
Bacillus macerans
RLM-013C
ATCC 202135
The species of each of these Bacillus bacterial strains were determined using the Fatty Acid Methyl Ester (FAME) Analysis method performed by Microcheck, Inc., of Northfields, Vt. using Trypticase Soy Broth Agar (“TSBA”) version 3.9 at 28.0 C of Microcheck's database. Strain RLM-013C, deposited as ATCC 202135, was determined by the “FAME” analysis to be
Paenibacillus macerans
GC subgroup A, which is also sometimes referred to as
Bacillus macerans.
The eight novel Bacillus strains have a variety of potential uses, including treatment of wastewater, such as municipal, food processing, and residential wastewater; pretreatment of wastewater that might contain high levels of oleaginous matter; treatment of on site sewage disposal systems, such as septic tanks and cesspools; agricultural uses, including livestock manure waste management or as Direct Fed Microbials for use in livestock feeds to increase food digestion and utilization; and for treatment of pipes, traps, plumbing systems and grease traps.


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