Enzymes are special types of proteins that act as catalysts to biological reactions.

Compared to
inorganic cataysts, enzymes can lower the activation energy of reactions better. Common
industries that use enzymes are food and beverage, textile, detergent, and pharmaceuticals.

Enzymes have been widely used in several industries like food production and manufacture
of commodities. Proteases remain the dominant enzyme type, because of their extensive
usage in detergent and dairy products while celluloses and amylases are used in other
industries such as textile and baking. The utility of enzymes will increase in the chemical,
pulp and paper, textile, waste treatment, pharmaceutical and diagnostic industries. Other
specialty applications include the use of enzymes in analytical applications, flavor
production, protein modification, personal care products, DNA technology and fine chemical
production. In the Philippines, most of the local industries rely on imported enzymes for
basic, medical or industrial uses.
Detergent Industry Enzymes in this industry is treated as detergent additives. The major
component is proteases, but other and very different hydrolases are introduced to provide
various benefits, such as the efficient removal of specific stains. There are new enzymes
which are more improved than the usual; proteases and amylases are being developed.
Protease is for protein-stain removal while Amylase is for starch-stain removal. The most
recent introduction of a new enzyme class into a detergent has been the addition of a
mannanase the result of a joint development between Procter and Gamble and
Novozymes.
Starch Conversion A good example of starch conversion is the enzymatic conversion of
starch to high fructose. Consecutive use of several enzymes is necessary for this process.
Common example of enzyme used is -amylase. Engineering efforts have also been
undertaken to develop improved versions of the enzymes used later in the process (i.e.
glucoamylase and glucose isomerise).
Textile Applications Enzymes play a big role for textile production. Some enzymes that are
used are Cellulase for denim finishing and cotton softening, Amylase for de-sizing, Pectate
lyase for scouring, Catalase for bleach termination, Laccase for bleaching and Peroxidase
for excess dye removal. An alternative, enzyme-based process performed at much lower
temperatures and using less water has now been developed based on a pectate lyase.
Feed Industry - Xylanases and -glucanases are the common enzymes which function as
feed additives and help in digestibility. The most recent advancement in feed enzymes have
been aimed at the improvements of phytases. Better utilization of feed is achieved by using
phytases. There are other approaches for the development of more effective enzymes. An
example of improved enzyme is Aspergillus fumigates phytase.

Food Industry - Enzymes are applied to processed food products as processing agents
upstream from the final product. Protease is used for milk clotting, Lipase for cheese
flavour, Lactase for Lactose Removal, Pectin methyl esterase for Firming fruit-based
products and Pectinase for Fruit-based products are just some of the most commonly used
enzymes in food production. Several advances have been made in the optimization of
enzymes for existing applications and in the use of recombinant protein production to
produce enzymes that do not have side-effects.

Enzyme Application in the Textile Industry

Enzyme biotechnology for sustainable textiles
Abstract
Enzymes are used in a broad range of processes in the textile industry: scouring, bleach cleanup, desizing, denim abrasion and polishing. Enzymes are specifi c and fast in action and small
amounts of enzyme often save large amounts of raw materials, chemicals, energy and/or water.
This chapter describes enzyme use in the textile industry in the context of sustainable
production and reports life cycle assessments (LCAs) on two enzyme applications: bioscouring
and enzymatic bleach clean-up. The results show that resource use and impact on the
environment can be reduced considerably when enzymes are implemented in the two
processes.
Introduction
In many industries, enzymes are used as biological catalysts to replace harsh chemicals
or perform reaction under milder conditions. In textile industry, enzymes not only make good
economic sense by saving energy, water and chemicals by improving quality, they also give
valuable environmental benefits. These benefits are very important in time as there is an
increasing awareness in the issue of sustainable development and climate change.
Novozymes recently performed two life cycle assessment (LCA) studies at textile mills in
China, one of the prime producing countries for cotton and textiles. One of the studies was of a
process known as bioscouring for removing impurities from cotton yarn. This is an alternative to
traditional scouring that involves a number of high-temperature steps with a large consumption
of chemicals. The other process is known as bleach clean-up to remove excessive bleaching
agent prior to dyeing. This enzymatic process required less water and less energy than the
conventional process used in China. On the basis of a qualitative assessment, it might seem
obvious to assume that enzymes contribute to sustainable development. But what are the hard
facts about the environmental impact of the use of enzymes in the textile industry? It should not
be forgotten that the production of enzymes is also associated with environmental burdens
(Nielsen et al., 2007). The purpose of the LCA studies presented here is to assess and compare
the environmental burdens created by enzyme production and distribution in comparison

with the environmental burdens avoided in the processes at textile mills. Here are two concrete
examples from China based on the specific conditions at two mills. It could be argued that these
facts only apply to the two specific mills. Therefore sensitivity analyses have been performed to
look at a variety of scenarios, such as the use of fuels other than coal to generate electricity or
the use of further optimised processes. In all cases, enzymes gave clear environmental benefits
and helped to reduce contributions to global warming.
Objective
The objective of the study of the two processes is to compare the process without the
use of enzymes and with the use of enzymes. Life cycle assessment was used for the study.

Environmental assessment of the enzymatic scouring of package cotton yarn for

dark-shade dyeing as an alternative to conventional chemical scouring
The assessment took place at the Rongxin Fibre Co. Ltd yarn dyeing mill located in
Haining industrial park, 90 km from Shanghai in the Zhejiang province of eastern China. Here
conventional scouring was replaced by enzymatic scouring with Scourzyme 301 L in 2007 and
enzymatic scouring is now running in full-scale production.
Light-colour yarns where pre-bleaching is required constitute about 80% of the market
and dark shades constitute about 20%. The study addresses the 20% part of the market
because the potential of bioscouring is largest for this product group. The reason for this is that
pectate lyase does not remove seeds and other seed fragments, which is a problem when
dyeing in lighter shades unless a natural look is preferred. Conventional scouring and
bioscouring can be performed on the same production line with the same production equipment.
Therefore there is no significant capital investment associated with a switch between the two
scouring methods. The study refers to production at a full-scale commercial yarn dyeing line at
Rongxin yarn dyeing factory with an output of 50 000 kg of dyed yarn per day. Chemicals are
delivered mainly by local producers. Water used in production is extracted from a nearby river
and treated by means of filtration and softening. Steam is supplied by the coal-fired Haining
Dong Shan combined heat and power plant located about 1 km from the factory, and electricity
is obtained from the national grid. Heat is not recovered in the textile mill. Wastewater generated
in the process is initially treated on site and then fed to a central treatment plant before it is fed
into a nearby river.
Methodology
Scouring is performed in a bath where a liquor of water and chemicals/enzymes is
pumped through the yarn. The liquor is heated with steam to an appropriate temperature, and
pumped through the yarn with an electric pump. Water and chemicals/enzymes are mixed with
each other in a separate tank next to the yarn bath.
Conventional scouring was performed in three steps: a scouring process followed by two
rinsing processes. The scouring process was performed at 100 C with penetration agent,
sodium hydroxide (NaOH) and hydrogen peroxide (H2O2). The penetration agent is a surfactant
that facilitated the waters entry into the yarn. Sodium hydroxide removed the unwanted
impurities and hydrogen peroxide bleaches the yarn. Sodium hydroxide is not selective in the
substrates that it attacks. In addition to the unwanted impurities, it also degrades other
substances in the yarn, including the cellulose.

Removed impurities were washed out of the yarn in the two rinsing steps. The first
rinsing step was performed at 95 C. The second rinsing step was performed at 50 C and
acetic acid was added to reduce pH. The starting temperature of the two rinsing steps was
higher than the starting temperature of the scouring process because the wet yarn and the dye
machine contained a large amount of residual heat transferred from the high temperature
scouring step. The entire scouring process took 3 hours including filling and emptying the bath.

Figure 1 Process diagram for conventional scouring. Upward slopes indicate a rise in temperature.
The final downward arrows indicate emptying of the bath.

Bioscouring is performed in a single process at 60 C with Scourzyme 301 L and

penetration agent. Scourzyme 301 L is an enzyme product containing a pectate lyase that
degrades pectins into soluble compounds. No rinsing steps or neutralisation steps are required
prior to the yarn dyeing. The penetration agents used for bioscouring can be the same as used
in the conventional scouring process. Bioscouring takes 90 minutes including the fi lling and
emptying of the bath. Water from conventional scouring and bioscouring, as well as from any
rinsing steps, is discharged directly for wastewater treatment after use.

Figure 2 Process diagram for bioscouring

Results

The results of the environmental impact assessment are shown in Fig. 3. Figure 3 shows
that resource consumption and environmental impacts induced by enzyme production generally
are very small compared with the savings. The reason is that a small amount of enzyme saves
large amounts of chemicals, energy and water in the scouring process. Transport of the enzyme
from Denmark to China does not add significantly to the environmental impact of Scourzyme
301 L even though the transportation distance is rather long. The explanation is that the quantity
of enzyme used is small and that ocean freight is energy efficient.

Figure 3 Added and saved resource consumption and environmental impacts when switching from
conventional scouring to bioscouring. All data are per tonne of yarn (dry weight).

The main factors behind the saved contributions to global warming are shown in Fig. 4,
which demonstrates that the heat saving in the bioscouring process is the main factor behind
the reduced contribution to global warming, followed by electricity and yarn savings. Savings of
water and chemicals in the process and the transport of chemicals from the manufacturers to
Rongxin are less important. Reduced water consumption in the scouring process explains most
of the water savings in Fig. 3. Water savings resulting from the reduced consumption of
chemicals and yarn (primarily from the irrigation of cotton fields) are of minor importance. The
saving of agricultural land observed in Fig. 3 can be entirely explained by the reduced cotton
production needed as a result of the use of enzymatic scouring.

Figure 4 Main factors behind saved contributions to global warming.

Environmental assessment of enzymatic bleach clean-up of light-coloured

package yarn and knitted fabrics as an alternative to rinsing with hot water
Fabric and yarn need to be dyed evenly and reproducibly to meet the quality
requirements of customers. For this reason, fabrics and yarns intended for fi nal products in light
colours are often bleached prior to the dyeing process. Bleaching is usually performed with
hydrogen peroxide and the excess bleaching agent must be carefully removed prior to dyeing to
avoid breakdown of the dye during the dyeing process. Bleach removal has traditionally been
performed by rinsing in several baths with water or by adding a reducing agent such as sodium
thiosulphate. Both of these traditional processes use considerable amounts of water and energy
for heating the water. Enzymes are now used as an alternative by many textile mills that dye
yarn and fabrics. Enzymatic bleach clean-up is based on a catalase, which catalyses the
conversion of hydrogen peroxide into water and oxygen. The LCA study took place at a textile
mill in the Esquel Group located in Guangdong in South East China where bleach removal
based on rinsing with hot water was replaced by enzymatic bleach clean-up with Novozymes
Terminox Ultra 50 L in 2002. The enzymatic process has been running in full-scale operation
since then.
Methodology
Bleach clean-up of package yarn is performed in a series of batch processes in water
mixed with chemicals/enzymes at appropriate temperatures. Liquid is continuously pumped
through the yarn rolls or the knitted fabrics to establish the necessary contact with the cotton.

Baths are heated with steam, and the pumping of water is facilitated by electric pumps. The
liquor ratio is 10 tonnes of water per tonne of yarn.
Reference for bleach removal. The traditional way for the mill to remove bleach is a
three-step process composed of one hot rinse and two cold rinses with pH adjustment in the
second cold rinse. The output from the process is bleached yarn or fabric without the presence
of hydrogen peroxide. The yarn or fabric is then dyed in a new bath.

Figure 5 Process diagram for the traditional way of removing bleach by rinsing with hot water. Upward
slopes indicate a rise in temperature. The final downward arrows indicate emptying of the bath.

Enzymatic bleach clean-up. Enzymatic bleach clean-up is performed in a two-step

process composed of one cold rinse and a cold hydrogen peroxide decomposition step where
the enzyme removes the hydrogen peroxide. Acetic acid is used for pH adjustment. The enzyme
works at high speed and the bath is free of hydrogen peroxide after 25 minutes. Then dyeing is
performed in the same bath with the same water.

Figure 6 Process diagram for enzymatic bleach clean-up.

Results
The results of the environmental impact assessment are given in Fig. 7 which shows that
the consumption of resources and environmental impacts caused by enzyme production are
generally very small compared with the savings, except in the case of agricultural land use. The
explanation is that a small amount of enzyme saves large amounts of energy and water in the
bleach clean-up process. Just as in the case of scouring with enzymes, transport of the
enzymes from the enzyme factory in Denmark to China does not add significantly to the
environmental impact of using enzymes.

Figure 7 Added and saved resource consumptions and environmental impacts when switching from the
traditional removal of bleach based on rinsing with hot water to enzymatic bleach clean-up. All data are per
tonne of package yarn or knitted fabric (dry weight).

The main factors behind the saved contributions to global warming are indicated in Fig. 8
which shows that the heat saving is by far the most important factor behind a reduced
contribution to global warming.

Figure 8 Main factors behind saved contributions to global warming.

Toxicity has not been included in the quantitative assessment for the same reasons as
explained for scouring. For the same reasons as before, it is considered very likely that the
toxicity impact reflects the other impact categories in Fig. 7 and that the use of enzymes
reduces the toxicity impact on the environment.

Conclusions about the two studies

The use of enzymes in scouring and bleach clean-up as alternatives to chemical
treatment and rinsing with hot water, respectively, led to considerable environmental
improvements at the two production lines at textile mills in China. The explanation is that a small
amount of enzyme saves considerable amounts of energy and water in both cases and also
chemicals in the case of scouring. Sensitivity analyses indicate that the general conclusion of
the assessment holds up under different energy supply scenarios although the sizes of the
reductions in environmental impacts are subject to much variation and uncertainty. The impact
of the transport of enzymes from the manufacturer to the final user is insignificant even though
the transportation distance is long. The main findings of the study are therefore applicable to
other textile mills with similar production systems elsewhere in the world.
The magnitude of the environmental improvements obtained by replacing the existing
production methods with the enzymatic technologies are highly dependent on the type of fuel
used and the actual production conditions. An estimation of environmental improvements at
other factories must therefore rely on specific information on production processes and energy
supply systems. The study has not addressed the removal of bleach with a reducing agent and
further environmental assessments are required before any conclusions can be made about this
method.

Silk Industry
History of silk

Sericulture or silk production has a long and colorful history unknown to most people. For
centuries the West knew very little about silk and the people who made it. For more than
2000 years the Chinese kept their secret and was the most zealously guarded secret in
history. History of silk began in the 27 century BCE. The key to understanding the great
mystery and magic of silk, lies with one species: the blind, flightless moth, Bombyx mori. It
lays 500 or more eggs in four to six days and dies soon after. From one once of eggs come
about 30,000 worms which eat a tom of mulberry leaves and produce twelve pounds of raw
silk, but this silk worm is unique in China.
th

Production
Producing silk is a lengthy process and demands constant close attention. To produce high
quality silk, there are two conditions which need to be fulfilled preventing the moth from
hatching out and perfecting the diet on which the silkworms should feed. Chinese
developed secret ways for both.
For the commercial production of silk silkworms are cultivated and fed with mulberry
leaves. The eggs are hatched by artificial means such as an incubator, and in the olden
times, the people carried it close to their bodies so that it would remain warm. Silkworms
that feed on smaller, domestic tree leaves produce the finer silk, while the coarser silk is
produced by silkworms that have fed on oak leaves. From the time they hatch to the time
they start to spin cocoons, they are very carefully tended to. Noise is believed to affect the
process, thus the cultivators try not to startle the silkworms.
Once the cocoons are ready then cultivators gather the cocoons and the chrysales are
killed by heating and drying the cocoons. In the olden days, they were packed with leaves
and salt in a jar, and then buried in the ground, or else other insects might bite
holes in it. Modern machines and modern methods can be used to produce silk but the
old-fashioned hand-reels and looms can also produce equally beautiful silk. In the process
of silk many steps runs in cascade manner in order to have a complete process of silk
production. Many process like removal of raw silk, washing, removal of impurities and dying
and drying are bit essential but removal of wax and other unwanted protein form raw silk is
most important step which make difference in the normal and quality silk known as
degumming.
Silk today
World silk production has approximately doubled during the last 30 years in spite of manmade fibers replacing silk for some uses. China and Japan during this period have been the
two main producers, together manufacturing more than 50% of the world production each
year. During the last 1970s China, the country that first developed sericulture thousand
years ago dramatically increased its silk production and has again become the worlds
leading producer of silk.
Silk industry in the Philippines
At present, the development of the silk industry is spearheaded by the Fiber Industry
Development Authority (FIDA), with the joint cooperation of PTRI, the Sericulture Research
and Development Institute (SRDI) of the Don Mariano Marcos Memorial State University
(DMMMSU), the University of the Philippines at Los Baos (UPLB) and other state
universities and colleges (SUCs).

The Organization for Industrial, Spiritual and Cultural Advancement-International started in

1999 by R. Shigemi Watanabe. Initially, the project is implemented to improve the economic
condition of the local farmers who are dependent on the declining sugar industry. It is
designed to create a source of livelihood among the locals living in mountainous areas
through the cocoon production and secure a stable source of income for the women in rural
areas through weaving and/or manual extraction of spun silk from the silk waste. It is found
in Negros Occidental and Negros Oriental
Degumming
Silk degumming process is a fundamental finishing process for silk yarn and silk fabric. The
objective of degumming is remove the substrate such as silk gum (sericin), wax and some
impurities from silk fibers. The principle of degumming process is breaking the peptide
linkage of amino acid in sericin structure into a small molecule, which is soluble in water.
The methods used for the degumming are the hydrolysis reaction performed by acid
and alkaline, but these methods are not eco-friendly and same time they have a big
problem on the surface area of silk. Hence the proteolytic enzyme have been used to solve
this problem and shown promising results not only in the production level but also quality of
silk. Since protease based degumming is eco-friendly which will be an additional advantage.
Though the conventional protease are quite efficient for degumming but having some
disadvantage like thermal and chemical stability which is one drawback has to solved, also
alkaline protease can hamper the quality and physical appearance of silk as silk is quite
sensitive to alkali and alkaline protease. The thermostable protease basically forms
Geobacillus genus has been used for the enzymatic degumming of silk which are quite
resistant to various chemicals and temperature. In this article we have summarized
comparative analysis of detergent, conventional protease and thermostable protease for
efficient degumming.
The principle behind silk degumming is increasing the silk gum solubility by breaking the
peptide linkages of sericin structure into small molecule such as amino acids and its
oligomer with hydrolysis reaction. Silk degumming can be performed by numerous methods
such as using alkaline and synthetic detergent, though it can be harmful to the silk because
of silks poor resistance to alkaline. In present the proteolytic enzyme is used to solve this
problem, though it has disadvantages and high costs.
Two process have emerged in the recent years, H.T.-HP. Degumming and Enzyme
Degumming. The High Temperature-High Pressure degumming requires special pressured
equipment and is energy intensive process. The Enzymatic degumming is emerging as ecofriendly fiber-gentle process where proteolytic enzymes that are effective under alkaline,
neutral as well as acidic conditions are being used.
Soap Degumming
Soap has been used in the process of silk degumming with synthetic detergents implies the
partial or total replacement of soap with synthetic non-ionic surfactants. To achieve the
better yield and quality soaps have been combine with an oxidizing or reducing bleaching
and, in some cases, even with dyeing, thus improving water and energy saving. Generally,
alkali and detergent mixtures are used at temperatures around 95 C - 98 C. Such a
treatment is suited to continuous processing.
The detergent based silk degumming leads to the production of low quality of silk
as detergent interfere with the mechanical properties of silk fibers. The fibroin, main part of

silk protein provides tensile strength to silk fibers which is very sensitive to soap,
alkali and alkaline protease. Another complication with detergent based silk degumming as
chemicals used for process are not eco-friendly hence long term effects on the surrounding.
Generally in the detergent based silk degumming soap derived from higher fatty acid has
been used which still remains in the silk even after many washing.
Microbial protease in silk production
The conventional detergents are not the choice for silk degumming as these chemical will
interfere the basic physical and chemical properties of silk leading to the complication in the
quality of silk. Though chemical based detergents have been used for long time but same
time silk produced by these methods always has shorter shelf life and various other
complications like less tensile strength, more hygroscopic etc.
Proteases cover the 60% of total enzyme market and amongst the most valuable
commercial enzyme. Alkaline proteases hold a great potential for application in the
detergent and leather industries and there is an ever increasing trend to develop
environment friendly technologies. Plants, animals and microbes are the main sources for
protease production. The preferred sources of proteases are microbes because of their
rapid growth and the ease with which they can be genetically manipulated to generate
new enzymes with altered properties and are currently being utilized by the
detergent industry eg. Serine proteases produced by Bacillus strains. Proteases from
several bacteria have been purified and characterized. Genus Pseudomonas a gramnegative bacterium that predominantly produces alkaline proteolytic enzymes and the
proteases has been purified. Fungal alkaline proteases are advantageous because of the
ease of downstream processing to prepare a microbe-free enzyme at low cost
production.
The protease and more specifically alkaline protease have shown numerous complication
with the silk during the process of degumming. First of all the silk is highly sensitive to the
alkali and alkaline protease which hampers it physical appearance. The Mechanical
properties also get affected under the exposure of alkaline protease. Another
complication with the conventional protease is their range of activity and stability in
various pH and Temperature. Another significant complication especially in protease based
silk degumming incorporation of chemicals in certain instants which also inactivates
the conventional enzymes.
Thermo-stable Protease
The optimal activity of conventional protease ranges 30-40`C which is not appropriate for
compete degumming process. Even some time process has to run for more than 24 hour in
at lower temperature which again causes deactivation of conventional enzyme. Often the
degumming process runs at higher temperature which leads to denaturation of conventional
proteases which are not thermo-stable. In some cases during the process a lots of chemical
are required to complete the process and many circumstances presence of chemical also
leads to the deactivation of enzyme and process subsequently.
The proteases are generally classified into two broad categories (exopeptidases, that
cleave off amino acids from the ends of the protein chain and endopeptidases, which cleave
peptide bonds within the protein) are becoming major industrial enzymes, and constitute

more than 65% of the world market. These protease enzymes have been extensively
used in the food, pharmaceutical, leather and textile industries.
The Bacilli provides 70% of protease hence the diverse sources has made these organisms
the focus of attention in biotechnology. In a chemically defined medium, thermophilic and
alkaliphilic Bacillus sp. JB-99 was also reported to produce thermostable alkaline
proteases. Dominant producers of proteases in fact, are the microorganisms of the
genera Pyrococcus, Thermococcus and Staphylothermus. Extremely thermostable serine
proteases are produced by the hyperthermophilicarc haeum Desulfurococcus strain, and
thermostable metalloroteases are reported from a gram-negative thermophilic bacterium.
In the last decades numbers of commercial uses have been established for
thermostable amylases and roteases. In the brewing industry, starchy materials used as
adjuncts must first be liquefied by the addition of thermostable amylases. Thermostable
amylases are used in desizing textiles to remove unwanted starch. They also may be used
to aid in the clarification of fruit juices in the manufacture of jelly and chocolate syrups.
Proteases are used in the leather industry, particularly in baiting hides to remove unwanted
inter fibrillar material.
Protease based silk degumming has been shown higher efficiency, expressed over removal
of sericin and energy save, achieved through lower process temperature. Milder
treatment conditions under which the fibers were processed during enzymatic
degumming prevented fibrillation and dusting i.e. fiber damage. Damage to the soap
degummed fibers was enhanced in subsequent dyeing process, which was the reason
of inability to spectrophotometrically measure dyestuff concentration in dye-bath. When
dyeing silk fibers, method of degumming should be considered. After the degumming fibers
still had lustrous, soft and smooth surface. Result of staining test with direct dyes was
appeared pink, indicating that there was a small amount of sericin remaining. This process
can be used instead of conventional degumming method with low cost and do not harm the
environment.
References:
1. Ole Kirk*, Torben Vedel Borchert and Claus Crone Fuglsang: Industrial enzyme
applications. Current Opinion in Biotechnology 2002, 13:345351
2. http://biotech.uplb.edu.ph/index.php/en/research3/food-feeds-and-specialtyproducts-biotechnology?catid=35:static&id=52:enzyme-research-lab
3. Silkroad Foundation. 2000. http://www.silk-road.com/artl/silkhistory.shtml . Accessed
September 23, 2014.
4. Republic of the Philippines, Department of Agriculture. Fiber Industry Development
Authority. http://fida.da.gov.ph/Templates/silk_history.htm . Accessed September 23,
2014.
5. OISCA. 2014. http://www.oisca-international.org/programs/sustainable-communitydevelopment-program/philippines/development-of-sustainable-communities-innegros-province-through-silk-production-/ . Accessed September 23, 2014.
6. Rajasekhar, A., Ravi, V., Reddy, M. N., & Rao, K. R. S. S. (2011). Thermostable
Bacterial Protease - A New Way for Quality Silk Production, 3(4), 4358.
7. P. H. NIELSEN, H. KUILDERD, W. ZHOU and X. LU, Novozymes A/S, Denmark

8.