>Woopex_oo1 wrote:
>Two questions:
>1. How do you culture the enzyme-producing fungus without using up
>too much of the cellulose feedstock that is supposed to make ethanol?
Answer: In general is better to buy a well known fungus from American Type
Culture Collection or from and industrial strain from some Biotechnoly
companies and grow it in small o large scale with an broth with K+, Ca++,
Mg, Po4, SO4 =, NO3 -, Fe +++, micronutrients and some cotton fiber or
paper pulp. Mainly the fungi with enzymes for cellulase and hemicellulase
will be selectively developed, excreting their enzymes in the broth to uso
the cellulose and hemicellulose as carbon source. If there is no
contamination, filter the fungi mycelia with sterilized paper filter and
then 0.5 micron sterile filter. Now you have a crude enzyme. If you want to
separate the salts, it could be done with ion resin, or you may precipitate
some enzymes using some salts, like amonium sulfate, test first, then
separate the presipited enzyme by filtration.
Use the crude enzyme or the presipited enzyme on your finely ground and
clean cellulose to get some fermentable sugars.
Look for the right yeast that could tolerate this medium to produce
ethanol.
If you go trying to isolate some fungi from wilderness take good care, some
might produce lung infections if your inmune system is not in good shape.
In an university library, in many biotechnoly journals you will find lots
of information and not only with fungi but reconbinant bacteria as well.
See: Dien et al., "Conversion of Corn Milling Fibrous Co-products
into Ethanol by
Recombinant Escherichia coli Strains K011 and SL40," World Journal of
Microbiology &
Biotechnology, 13, 619-625 (1997).
Leathers et al., "Saccrification of Corn Fiber Using Enzymes from
Aureobasidium sp. Strain
NRRL Y-2311-1," Applied Biochemistry and Biotechnology, 59, 337-347
(1996).
There are many ways to repare these array of enzymes Hemicellulase,
Cellulase; I search
some lates Patent on this issue:
U.S. Patent and Trademark Office
http://www.uspto.gov/patft/ > Patent Number Search
United States Patent 6,423,524 Hagen, et al. July 23, 2002
Cellulase preparation comprising an endoglucanase enzyme
Abstract
The present invention relates to cellulase preparations consisting
essentially of a
homogeneous endoglucanase component. The cellulase preparation may be
employed in the treatment of cellulose-containing fabrics for
harshness reduction,
for color clarification, or to provide a localized variation in the
color of such fabrics,
or in the treatment of paper pulp.
>2. What is the enzyme that breaks down lignin? And which fungus
>produces it?
Answer: From the word lignine the enzyme is called Ligninase, and lignins
are composed by may complex phenol derivative compounds and we usually want
to get rid of lignins by oxidazing them, that is another way to call the
enzymes are phenol oxidizing enzymes. Some industial use of these enzymes
are disclosed for textile bleaching in:
U.S. Patent and Trademark Office
http://www.uspto.gov/patft/ > Patent Number Search
United States Patent 6,384,007 Convents, et al. May 7, 2002
Method and composition for enhancing the activity of an enzyme
Abstract
There is provided a process for enhancing the activity of a phenol
oxidizing enzyme, comprising adding to the enzyme, as an enhancer for the
activity of said enzyme, one or more compounds having the having the
formula:
##STR1## wherein Z.sub.1 and Z.sub.2 are electron withdrawing groups,
independently selected from the group consisting of optionally substituted
alkyl/(hetero)aryl- -sulfone, -sulfoxide, -sulfonate, -carbonyl, -oxalyl,
-amidoxalyl, -hydrazidoxalyl, -carboxyl and esters and salts thereof,
-amidyl, -hydrazidyl, nitrile. The process is especially useful for
removing colored stains from fabrics in a washing process.
------------------------------------------------------------------------
-----------------
United States Patent 6,242,245 Amann, et al. June 5, 2001
Multicomponent system for modifying, degrading or bleaching lignin or
lignin-containing materials, and processes for its use
Abstract
A multicomponent system for modifying, degrading or bleaching lignin and
lignin-containing materials or similar substances, includes an
oxidoreductase and an oxidant suitable for the oxidoreductase and a
mediator and at least one enzymatically active additive. The mediator does
not inactivate the oxidoreductase and the enzymatically active additive,
and the enzymatically active additive is selected from the group consisting
of the hydrolases of the enzyme class 3.2.1.
------------------------------------------------------------------------
-------
United States Patent 6,426,410 Wang July 30, 2002
Phenol oxidizing enzymes
Abstract
Disclosed herein are novel phenol oxidizing enzymes naturally-produced by
strains of the species Stachybotrys which possess a pH optima in the
alkaline range and which are useful in modifying the color associated with
dyes and colored compounds, as well as in anti-dye transfer applications.
Also disclosed herein are biologically-pure cultures of strains of the
genus Stachybotrys, designated herein Stachybotrys parvispora MUCL 38996
and Stachybotrys chartarum MUCL 38898, which are capable of
naturally-producing the novel phenol oxidizing enzymes. Disclosed herein is
the amino acid and nucleic acid sequence for Stachybotrys phenol oxidizing
enzymes as well as expression vectors and host cells comprising the nucleic
acid. Disclosed herein are methods for producing the phenol oxidizing
enzyme as well as methods for constructing expression hosts.
Best regards
Juan
--- In [EMAIL PROTECTED], Juan Boveda <[EMAIL PROTECTED]> wrote:
> The method to convert hole plants composed by cellulose,
hemicellulose and
> lignins uses ezimes and water to hydrolyse these material to
soluble
> sugars, theses enzimes are true catalysers coming from mainly fungi
and
> some bacteria. These enzimes are called cellulase and hemicellulase.
>
> Many fungi has been isolated and are used by biotechnology
>companies to produce cellulases in large scale. One of the
>common uses of cellulases is
> to finish blue jeans with a soft "stoned washed" touch.
>
> Some fungi cellulase (the specific enzime for cellulose) come from
> Aspergillus niger, Trichoderma viride, Penicillium funiculosum, etc.
> they usually are a mixture of enzimes with different activities or rate
> of conversion of cellulose to fermentable sugars in a given time.The
> hemicellulases are produced as well by many of these strains for
example
> Aspergillus niger.
>
> The hardest to degrade are the lignins, few fungi are able to do it
soo,
> because lignins are toxic compouds to most of them.
>
> Many of these fungal enzimes has maximun activities on mild acid pH
4 to 7
> and mild temperatures 20 - 50o C.
>
> Regards,
>
> Juan
> Woopex_oo1 wrote:
>
> -----Mensaje original-----
> De: womplex_oo1 [SMTP:[EMAIL PROTECTED]
> Enviado el: Jueves 29 de Agosto de 2002 12:48 PM
> Para: [EMAIL PROTECTED]
> Asunto: [biofuel] Cellulose - to - Sugar Preprocessing
>
> What energy efficient, eco-friendly methods exist to convert
> cellulose to sugar so that entire plants - leaves, stem, roots &
all -
> can be fermented into ethanol? I've heard termites do this
> routinely...
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