Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Fungi
Reexamination Certificate
1995-05-22
2002-04-30
Sherrer, Curtis E. (Department: 1761)
Chemistry: molecular biology and microbiology
Micro-organism, per se ; compositions thereof; proces of...
Fungi
C435S278000
Reexamination Certificate
active
06379948
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to a novel strain, and to a pulp bleaching method using the same. In particular, it relates to a novel lignin-degrading strain having high lignin degrading activity and pulp bleaching activity, and to a pulp bleaching method using the same.
BACKGROUND OF THE INVENTION
Conventionally, in the bleaching of wood pulp, chlorine and chlorine compounds were widely used to remove residual lignin in the pulp.
In recent years, however, it has become clear that these chlorine compounds generate dioxin, which is a toxic substance, and hence it is now necessary to limit their use as a matter of urgency.
Various bleaching methods have been proposed to replace chlorine compounds, and one such effective technique, “bio-bleaching” that makes use of strains or their enzymes, is now attracting considerable attention.
In the past, many attempts have been made to degrade the lignin in wood using strains for use in the paper pulp industry, etc.
A series of Basidiomycetes known as white rotted fungi that selectively degrade lignin were used for this purpose.
Coriolus versicolor and Phanerocheate chrysporium are typical examples of such fungi with lignin. degrading activity, and they have also been used in research and development.
However, these strain of fungi do not have sufficient lignin degrading activity, and they were therefore inadequate for improving the brightness of pulp. Moreover, their selectivity is low so that in addition to lignin, they degrade polysaccharides such as cellulose and hemicellulose at the same time. This leads to poorer pulp quality and lower pulp yields, hence it was so far impossible to use these fungi industrially.
The inventors studied methods of isolating and screening novel strain that had excellent pulp bleaching activity, and could be used industrially. They were thus able to develop such a method for novel strain that can identify and isolate lignin-degrading strain with high accuracy (Tokugan Hei 5-85731).
Based on the aforesaid selection method for lignin-degrading strain, the inventors carried out detailed studies, using rotted wood or fruiting body as a isolating source, in order to isolate novel strain having a high pulp bleaching activity, As a result, they succeeded in isolating a novel strain that not only has high lignin-degrading activity, but which is also capable of greatly improving the brightness of pulp, and thereby arrived at the present invention.
It is therefore a first object of this invention to provide a lignin-degrading strain for pulp bleaching that has both high lignin-degrading activity, and thermophilic pulp bleaching activity.
It is a second object of this invention to provide a pulp bleaching method that can be used industrially, employing this novel strain having high lignin-degrading activity and/or a culture prepared from the strain.
DISCLOSURE OF THE INVENTION
The aforesaid objects of the invention are achieved by the strain SKB-1152 that has high lignin-degrading activity and thermophilic pulp bleaching activity, and by a pulp bleaching method using this strain.
This invention will now be described in more detail. The novel strain SKB-1152 of this invention has the following microbiological characteristics:
(1) Growth in Culture Medium
Type of culture medium
Growth
Malt extract agar medium
+++
Potato extract dextrose medium
+++
Czapeck agar medium
+++
Sabouraud agar medium
+++
Synthetic mucor agar medium
+++
YpSs agar medium
+++
Culture conditions: 30° C., 3 days
+++
Growth: Slight +/Medium ++/Vigorous +++
+++
(2) Physiological and Ecological Characteristics
The pH required for growth (Potato extract dextrose medium, 30° C., 3 days culture) is in the range 3-9.
The strain will not grow at pH 2 or lower, or at pH 10 or higher. The optimum pH range is 4-8.
The temperature required for growth (Potato extract dextrose agar medium, pH 5.6, 4 days culture) lies in the range 20-40° C. The strain will not grow at 50° C. or higher.
The optimum temperature range is 28-37° C.
The phenoloxidase reaction (30° C., 3 day culture) gives a slight or negative reaction.
The colonies (Potato extract dextrose agar medium, pH 5.6, 4 day culture) resemble thin white felt.
Physiological and ecological characteristics, and mycological characteristics such as growth on various culture media, were examined in detail, however this strain could not be identified with any known strain.
As will be clear from the examples described hereinafter, this strain has better lignin-degrading activity and pulp bleaching activity than Coriolus versicolor and Phanerocheate chrysporium which have conventionally been regarded as typical lignin-degrading strain.
No conventionally known strain has both excellent lignin-degrading and pulp bleaching activity. The inventors therefore identified this fungi as a new strain, gave it the name SKB-1152, and deposited a sample to the National Institute of Bioscience and Human-Technology, Agency of Industrial Science & Technology, Ministry of International Trade and Industry (Contribution No. FERM P-4718).
As there are still some unresolved questions regarding the classification of this strain of fungi, a precise identification is difficult to make at this time and the results of future research must be awaited.
An outline of how SKB-1152 is isolated and selected will now be given.
According to this method, a culture medium containing lignin or pulp with residual lignin, is inoculated with a isolating source sample of rotted wood or fruiting body, and strain having lignin-degrading activity are isolated depending on the formation or non-formation of a decolorized area in the medium.
The lignin used herein may be Kraft lignin or sulfate lignin, and the pulp with residual lignin may be a chemical pulp such as Kraft pulp. The medium used for the culture may be a liquid medium wherein a culture solution is merely added to pulp, or a solid culture wherein pulp is solidified with agar or the like.
The preparation of the medium for the aforesaid isolation and selection method, and the selection of lignin-degrading strain, may be carried out according to methods known in the report.
This method of isolating and selecting lignin-degrading strain is simpler and more precise than the conventional method of phenoloxidase detection, and it also has an advantage in that lignin-degrading strain can be isolated and selected from rotted wood or fruiting body.
Next, the pulp bleaching method using the novel strain SKB-1152 of this invention will be described.
In the pulp bleaching method according to this invention, pulp is basically treated with SKB-1152 or a culture produced using this strain, and the strain is then cultured for a predetermined time. There is no particular limitation on the culture conditions provided that they are within the range of physiological characteristics of the aforesaid strain.
Herein, what is meant by a culture produced using SKB-1152 is a mixture of microbes and culture solution that has been obtained by culture of this strain. In the context of the invention, this includes not only microbes isolated from a microbial culture, but also the residues remaining after the microbe is isolated, and the culture solution remaining after all solid matter has been eliminated from the culture.
The pulp used in this invention may be a chemical pulp or mechanical pulp, there being no particular limitation on the type of pulp provided that it contains residual lignin. Moreover, there is no limitation on the form of the pulp used for the culture provided that strain grow easily on it and it is easy to bleach.
The pulp concentration, temperature and pH used for the culture may be chosen suitably within a range wherein the strain has physiological activity.
However, from the viewpoint of airation and growth of the strain, in the case of a static clture, the pulp concentration is preferably within the range 15-60%, while in the case of a shaking c
Iimori Takeshi
Kaneko Reiji
Machida Makoto
Yoshikawa Hiroshi
Nippon Seishi Kabushiki Kaisha
Sherrer Curtis E.
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