Beer for the ladies making great-tasting light beer possible

no. 03 september 2008 the quarterly bioindustrial magazine from novozymes

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Meeting customers close to home 4

South African bakeries fight rising costs with enzymes 6

Making a better soy sauce 8

Vietnamese soy sauce a step ahead of the rest 9

Half-life extension of drugs by design 10

novozymes is the world leader in bioinnovation. together

with customers across a broad array of industries we

create tomorrow’s industrial biosolutions, improving our

customers’ business and the use of our planet’s resources.

Pub lished by No vozymes a/s

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g e n e r a l

Many companies small and large are inter-

ested in becoming more sustainable and

the American retailer Wal-Mart is one of

them. Wal-Mart is the world’s largest retail-

er with sales of over USD 380 billion, 2 mil-

lion employees, and 60,000 suppliers. Wal-

Mart has strong and simple environmental

goals: to be supplied 100% by renewable

energy, to create zero waste, and to sell

products that sustain the environment.

Novozymes is doing its part in helping Wal-

Mart achieve its ambitious goals.

In August, Novozymes was honored to

be invited as the keynote speaker at Wal-

Mart’s supplier conference in Bentonville,

Arkansas, which focused on how to create

more sustainable products. Novozymes

presented its own experiences with sustain-

able solutions and contributed its ideas

on designing everyday consumer products

better.

rethinking tomorrow’s productsOf the countless goods that Wal-Mart sells

every day, many can be improved, and with

less environmental impact, using enzymes.

When enzymes are applied in the manu-

facturing of many of the products that

we use in our daily lives, chemical use can

be reduced, and water, energy, and other

resources can be saved.

Detergents, bread, textiles, cooking oils,

writing paper, and many other consumer

products can benefit from enzyme technol-

ogy. Microbial solutions also contribute to a

better environment and can directly replace

many of the chemical solutions used in

consumers’ homes, such as carpet cleaners

and bathroom solvents.

By inviting Novozymes to play a key role

in its supplier conference, Wal-Mart has

recognized Novozymes for its high sustain-

ability standard and expertise, and signals

that it considers enzymes and microorgan-

isms inherently sustainable solutions.

the biggest differenceEnzyme technologies often create the

potential for large energy savings for

manufacturers. It is therefore important

to inform corporate decision-makers, like

Wal-Mart and its suppliers, as well as public

opinion-makers that enzyme and microbial

technologies can play a very active part in

meeting the climate challenges.

“The biggest difference Novozymes can

make in reducing global CO2 emissions

is by contributing to the increased use of

enzyme and microbial technology,” says

Claus Stig Pedersen, head of Sustainability

Development at Novozymes. He explains

that in 2007 alone Novozymes’ products

reduced CO2 emissions by more than 20

million tons. Roughly estimated, this means

that the world saves 100 kg of CO2 emis-

sions when 1 kg of Novozymes’ enzymes is

used.

TAlkINg sustaiNaBility WITH WAl-MART

SUSTAINABlE RECOgNITION

Novozymes’ sustainability efforts and

solutions have not gone unnoticed. The

Dow Jones Sustainability group Index

has, for the 7th year in a row, selected

Novozymes for the #1 position in the

biotech sector. The United Nations global

Compact is the largest sustainability

initiative in the world with over 3,000

participating companies. For the 2nd year

in a row global Compact has recognized

Novozymes for its efforts to integrate sus-

tainability in its way of doing business.

b r e w i n g

Attenuzyme Flex is Novozymes’ solution to a

rising demand for light beers. The fight against

obesity has made consumers more conscious

about their calorie intake, and they often turn

to light and low-carb products whenever taste

and appearance are not affected. In the US,

half of the beer sold is light, and the market for

light and low-calorie beer is expected to grow

faster than the traditional beer market.

a new tool for brewmastersAs the latest addition to Novozymes’ portfolio of

enzymes for beer production, Attenuzyme Flex

has been highly welcomed in breweries around

the world. As in any other production, breweries

are always in search of effective process control

and improved economy, and enzymes are a

natural solution to both of these issues.

“Compared to the first generation of

Attenuzyme, Attenuzyme Flex is an optimized

blend of enzyme activities that works syner-

gistically with the naturally occurring amylases

in the malt,” says Ricardo gerlack, Customer

Solutions Brewing Scientist at Novozymes. “The

combination of these enzyme activities provides

a rapid and effective breakdown of the avail-

able starch into fermentable sugars.”

Attenuzyme Flex is an all-in-one, single-

dosage product containing a unique, patented

pullulanase component that ensures a faster

process and a more consistent performance.

And best of all, Attenuzyme Flex performs

optimally under standard mashing conditions

and does not require any changes in the

process.

time and energy savingsProducing highly attenuated beer requires

longer mashing times, which can decrease

brewhouse capacity. Attenuzyme Flex makes it

possible to shorten mashing times, providing an

improved utilization of the brewhouse capacity.

When added at the beginning of the main

mash, Attenuzyme Flex can shorten conversion

time by up to 50% while still maintaining the

desired attenuation. The optimized amount of

enzyme used depends on the desired degree of

attenuation, the mashing time, and the tem-

perature. The shorter conversion makes it pos-

sible to produce a highly attenuated beer in the

same amount of time as a standard beer. This

results in a faster process, increased brewhouse

capacity, and significant savings of both time

and energy.

gOOD NEWS FOR BEER lOVERS AND BEER MAkERS

Across the globe, low-calorie brews are gaining ground on

traditional beers. It is safe to say that this is not just a trend

– light beer is here to stay. Attenuzyme® Flex helps make

great-tasting light beer possible.

Attenuzyme® Flex

makes it easy to

produce great-

tasting light beer

that women enjoy.

3

b r e w i n g f u e l e t h a n o l

A sketch of the new

enzyme plant which

is planned to be

completed in 2010.

“Attenuzyme Flex allows breweries to realize a broad-

er range of wort attenuation, from highly attenuated

beers such as low-calorie and light beers to those requir-

ing only fine attenuation adjustments,” says Søren lund,

Regional Marketing Manager for Brewing at Novozymes.

“The sugar profile is more favorable with a lower

glucose- to-maltose ratio compared to wort produced

with regular glucoamylase-based enzymes, especially

for fine attenuation adjustments. A better sugar profile

means better yeast performance, which results in a more

desirable flavor profile.”

Attenuzyme Flex is inactivated during wort boiling and

is considered a processing aid. Furthermore, the reduc-

tion of enzyme load per brew also reduces problems with

wort filtration and hot break removal, which is often

seen with a highly saccharified mash.

Beer for the ladieslow-calorie and low-carb brews generally appeal to

women, and it will be interesting to see if these healthier

beers will inspire more women to choose a beer over a

mixed drink or glass of wine. Market experts find it likely

that new and more targeted marketing approaches will

focus on women in the years to come – a major change

in a market that has traditionally been solely directed

towards men.

Once again a Novozymes product contributes to the

greater good. Healthier beers with fewer calories and

reduced energy usage are the result when breweries

change to Attenuzyme Flex. And it is good to see that

consumers are ready for the change, too.

MEETINg CUSTOMERS ClOSE TO HOME

The new facility will be located on a 30-acre

property at the Biorefinery Campus in Blair,

ap proxi mately 25 miles north of Omaha,

Nebraska. Novozymes expects to break ground

in 2008 and start operations in late 2010.

the search begins“The search for the perfect location started 18

months ago,” says Per Olesen, Vice President and

project chairman at Novozymes. “Our objective

was to find the most economical and efficient

area from which to supply our American fuel

ethanol customers. To begin with we didn’t limit

Novozymes has selected Blair,

Nebraska, as the location for

its new USD 80–100 million

production facility. The plant

will produce enzymes for

existing corn-based fuel etha-

nol and later, enzymes for

cellulosic ethanol production.

“light beers are

here to stay, and

Attenuzyme® Flex can

help brewers meet

this consumer need,”

says Søren lund,

Regional Marketing

Manager for Brewing

at Novozymes.

for More iNforMatioNSøren lundshl@novozymes.com

4

for More iNforMatioNPer Olesenpo@novozymes.com

MEETINg CUSTOMERS ClOSE TO HOMEour options; we started looking across the whole

world for possible sites.”

The project team quickly narrowed the pos-

sibilities - first to the Americas, then the US, then

the Midwest, and finally to Blair, Nebraska.

“We found that proximity to our customers

means a great deal,” says Per Olesen. “There are

many excellent sites around the world to place a

new enzyme manufacturing facility. But when we

looked at the entire package, the Midwest was the

best match. There were many variables to consider,

including raw material sourcing, utilities supply,

transportation costs, construction costs, infrastruc-

ture, and the availability of a skilled workforce. In

Blair we identified a site where all of our needs

could be met in the most optimal way.”

enthusiasm all around“The US is an attractive and competitive market

for us to invest in,” explains Peder Holk Nielsen,

Executive Vice President at Novozymes. “The new

location will allow us to sustain our close coop-

eration with bioethanol customers across the

Midwest and provides us access to a well-

edu cated workforce and a good infrastructure.”

The state of Nebraska also warmly welcomes

the new facility. Nebraska State governor Dave

Heineman says, “Nebraska is pleased that

Novozymes has selected Blair for its world-class

biotech facility.”

Initially the facility is planned to bring approxi-

mately 100 new jobs to the state. Further devel-

opment could increase the number significantly.

David g. Brown, president and CEO of the

greater Omaha Chamber of Commerce, says,

“This is one of the most exciting investments

and job announcements we’ve had in recent

years.”

one piece of a larger planThe new facility in Nebraska is just one of the

important initiatives that Novozymes is taking to

grow with the American biofuel industry.

Earlier in 2008, one of the first steps was

taken to meet Midwest customers’ needs by

opening an office and laboratory facility in Ames,

Iowa, another city in the heart of the Corn Belt.

The main focus of the Ames office is to develop

strong relationships with Midwest customers and

provide quick-response technical support.

Expansion of both R&D facilities and produc-

tion is also taking place at Novozymes’ enzyme

facility in Franklinton, North Carolina.

Growing with the industryWith the American biofuel industry growing so

quickly, Novozymes wants to have the necessary

capacity to grow together with it. The expan-

sion initiatives being implemented right now are

key to preparing Novozymes for the inevitable

requirements of tomorrow’s biofuel industry - not

just in the US, but worldwide.

“When locating

a new production

facility, proximity

to your custom-

ers means a

great deal,” says

Per Olesen, Vice

President and

project chairman

at Novozymes.

5

C e r e a l f o o d

Bread is a staple food in South Africa but has

become much more expensive recently. The aver-

age price of wheat has doubled from March 2007

to March 2008, causing bread prices to increase

by over 20% in the same period. This is creating

tough times for the South African baking industry

and it is now looking for ways to cut costs while

maintaining quality. Enzymes from Novozymes

can help to minimize the impact of soaring costs.

Growing industrial segmentAbout 60% of all bread in South Africa is pro-

duced in small artisan bakeries and 40% by

industrial bakeries. Measured in value terms, sales

of industrially produced bread rose by 6% from

2006 to 2007, according to Euromonitor figures.

The share of the different types of bread made

by industrial bakeries in South Africa is shown in

Figure 1. Brown bread has gained in popularity in

recent years to account for 41% of sales whereas

white bread accounts for 51%.

The industrial bakeries are the primary tar-

get for South African companies making bread

improvers and bread mixes. They are increasingly

using enzymes from Novozymes in their formula-

tions.

savings compared to emulsifiersIn the last 12 months, the price of emulsifiers

has increased dramatically on the world market.

Increased prices for energy and for raw materials

such as tartaric acid are the main causes.

The use of emulsifiers such as DATEM and SSl

had been taken for granted by many bakeries in

South Africa, but with higher prices, the bakeries

are beginning to consider alternatives. lipopan

Xtra is a lipase enzyme that can be used to

replace emulsifiers either partially or completely.

It works well with the straight dough method

that predominates in South Africa. lipopan Xtra

enhances the authentic bread flavors while deliv-

ering good dough strengthening just as emulsi-

fiers do. lipopan Xtra currently gives a cost saving

of 40% to achieve a comparable performance.

Caking is a common problem with emulsifiers.

“In a hot, humid country like South Africa, the

emulsifier can form lumps that are as hard as

rock and must be broken up before use,” com-

ments Andre Diedrick, an Account Manager for

Cereal Food at Novozymes in South Africa. “Our

enzymes, on the other hand, are free-flowing

granulates.”

“You also save on storage space and logistics,”

he adds. “A typical dosage of lipopan Xtra is just

10 ppm of the dough. You can replace one ton of

emulsifiers with just 10 kg of enzymes.”

This small dosage makes many new customers

in South Africa skeptical toward lipopan Xtra at

first. They simply do not believe that such a small

amount of enzyme will have the desired effect.

That is why Andre Diedrick and his colleague

Vincent kibare perform trials in the Novozymes

baking laboratory in South Africa to prove the

robustness in industrial scale of lipopan Xtra.

“Slowly but surely we are changing the mindset

here, and we now have a wide range of different

customers for lipopan Xtra,” says Vincent kibare,

Account Manager.

fluctuating wheat qualityThe fundamental ingredient of bread – wheat

flour – has been in short supply on the world mar-

ket in 2007 and 2008, and this has also affected

costs for the baking industry in South Africa.

Vincent kibare comments: ”In general, the

poorer the quality of the wheat, the more you

need to use enzymes to produce good-quality

bread. An advantage of lipopan Xtra is that it

strengthens the dough regardless of the quality

of the flour. Even with a poor-quality wheat flour,

you still get good results.”

exorbitant ascorbic acidThe price of yet another basic bread ingredient

has also risen – ascorbic acid, which is used to

strengthen gluten. One kilogram of ascorbic acid

rose to the exorbitant price of ZAR 200 in South

Africa in June 2008, equivalent to about USD 25

per kilogram. This is a six-fold increase compared

to the price of only ZAR 30 per kilogram in June

2006.

China is a prime source of this baking ingredi-

ent, but some Chinese factories making ascorbic

acid have closed down recently due to strict pol-

lution control regulations being enforced. In addi-

tion to the baking industry, ascorbic acid (better

known as vitamin C) is purchased by the phar-

maceutical industry, which is able to pay a higher

price. This has diverted supplies to the pharma-

ceutical industry and forced the price up.

Novozymes can offer bakers an alternative

to ascorbic acid: glucose oxidase. The gluzyme

Mono product can be used as a partial replace-

ment for ascorbic acid to give the same results in

terms of dough stability (see Fig. 2). At current

prices, using gluzyme Mono is much more eco-

nomical than the equivalent amount of ascorbic

acid it replaces.

There are also additive effects from using both

gluzyme Mono and lipopan Xtra together, such

as increased loaf volume.

SOUTH AFRICAN BAkERIES FIgHT RISINg COSTS WITH ENZYMES

The combination of soaring prices for raw materials and a new local set-up for Novozymes has led

to increased sales of baking enzymes such as lipopan® Xtra and gluzyme® Mono in South Africa.

6

for More iNforMatioNVincent kibarevnck@novozymes.com Andre Allan Diedrickaadi@novozymes.com

the right platform for growthAlthough times are tough for the baking

industry, sales of Novozymes enzymes in South

Africa have actually performed very well.

A change of strategy at the end of 2006

has been instrumental for establishing the

right platform for Novozymes’ baking busi-

ness in South Africa. To serve the baking

industry in the best possible way, manage-

ment made the decision to concentrate

entirely on its primary business: developing,

producing, and selling pure enzyme solutions.

During the transition period in 2007,

Novozymes helped bakery customers to

secure new suppliers of products contain-

ing Novozymes enzymes. The changeover

has been well accepted by bakeries, and

Novozymes now supplies enzymes to bread

improver companies and premix manufactur-

ers, who in turn supply the bakeries.

Darrell Ward, Novozymes’ Industry Sales

Manager for Cereal Food in Africa among

other regions, has been spearheading the

implementation of the new strategy: “The

enzyme market in South Africa is looking

very promising for us with good growth in

the industrial baking sector. Much credit

goes to the local team of Account Managers

in South Africa.”

SOUTH AFRICAN BAkERIES FIgHT RISINg COSTS WITH ENZYMES

Andre Diedrick (left)

and Vincent kibare,

two Account Managers

for Cereal Food based

at Novozymes in South

Africa. As well as South

Africa, they cover other

countries in Africa

south of the Sahara.

Bread prices have risen

significantly in South

Africa as a result of

the increasing cost of

ingredients such as

emulsifiers and ascor-

bic acid, but enzymes

can be used to offset

the increases.

FIg. 2. EFFECT OF PARTIAl REPlACEMENT OF ASCORBIC ACID WITH glUZYME® MONO

BREAD TRIAlS USINg THE STRAIgHT DOUgH PROCESS

All loaves contain 5 ppm Fungamyl® 2500 Sg + 40 ppm Pentopan® Mono Bg. A dosage of 9 ppm

of gluzyme® Mono and 30 ppm ascorbic acid gives a result (far right loaf), in terms of loaf volume

and crumb structure, that is comparable to the loaf made using 90 ppm of ascorbic acid. At cur-

rent prices, the cost of using this enzyme is much less than the cost of the ascorbic acid it replaces.

30 ppm ascorbic acid 60 ppm ascorbic acid 90 ppm ascorbic acid 30 ppm ascorbic acid + 9 ppm gluzyme® Mono

FIg. 1. TYPES OF PACkAgED INDUSTRIAl BREAD SOlD IN SOUTH AFRICA IN 2007

White bread 51%

Brown bread 41%

Specialty bread 6%

Pita 1%

Tortilla 1%

Courtesy of Euromonitor

7

tured in a surface fermentation process for several

days. A salt solution is then added to create the

moromi, which is fermented to extract the flavor

of the soy and wheat. This mixture is further

fermented until the full flavor of the soy and

wheat is achieved. Then the mixture is filtered and

the raw soy sauce is extracted. Press cake and,

depending on the variety of soybean used, oil are

by-products of the process. The press cake is used

as livestock feed. The raw soy sauce is refined and

pasteurized before it is packaged.

Traditionally, the fermentation process for soy

sauce is time-consuming, often requiring many

months to infuse the correct flavor, aroma, and

color, as the natural fermentation progresses and

flavors are released by the enzymes produced dur-

ing this process. An alternative and much faster

solution is to predigest the soy and wheat with

enzymatic hydrolysis or acid hydrolysis.

Quicker alternativesPredigesting the main ingredients drastically

speeds up fermentation time, as it makes the

hydrolyzed peptides in the soy and wheat more

readily available for fermentation. Both enzymatic

and acid hydrolysis provide the high nitrogen con-

tent necessary for enhanced flavor formation.

The predigestion of soy and wheat takes place

f o o d & n u t r i t i o n

MAkINg A BETTER

SOY SAUCE

Enzymes can improve the soy sauce industry; infusing more flavor,

increasing yield, and reducing fermentation time. Enzymatic hydro-

lysis offers a simple process to ensure that the soy sauce tastes

great and at the same time is without dangerous levels of 3-MCPD.

There are almost as many ways to produce soy

sauce as there are countries in Asia. It seems

that most Asian countries have their own special

traditions regarding ingredients used, processing

practices, and fermentation times. In spite of the

many differences, enzymes can help manufactur-

ers produce tasty, healthy, high-quality soy sauce.

The main ingredients of soy sauce are soybeans

or defatted soybean meal, wheat or wheat bran,

salt, and water. The sauce is manufactured by a

complex fermentation process in which the carbo-

hydrates in the soy are fermented to alcohol and

lactic acid, and the proteins are broken down into

peptides and amino acids. This produces the typi-

cal brown color and distinctive soy sauce flavor

that is loved by millions of Asian food enthusiasts

around the world.

Enzyme technology can substantially improve

the industry, enhancing soy sauce flavor, increas-

ing yield by 5 to 8%, and reducing fermentation

time. The exact amounts of yield increase and

fermentation time reduction differ greatly from

manufacturer to manufacturer due to differences

in traditional practices and process equipment.

The extra yield is particularly likely in factories

that use presses to separate the moromi (mash)

into soy sauce and press cake. And since the

price of soy has skyrocketed during the last year,

the economic impact of increasing yields is much

greater than it had previously been.

traditional soy sauce productionAlthough the production of soy sauce varies

considerably, in most cases the three main ingre-

dients are soybeans, wheat, and salt. The wheat is

roasted and crushed to release its aroma, and the

soybeans are steamed until they are softened and

easier to process.

Typically, a starter (koji) consisting of the micro-

organisms Aspergillus oryzae and/or Aspergillus

sojae is added to the main ingredients and cul-

BeNefit aChieved

Higher yield in moromi separation

Reduction of fermentation time

More intense flavor profile

great-flavored HVP with insignificant 3-MCPD levels

eNzyMe(s) used

Viscozyme® / Celluclast®

Alcalase® 2.4 l Fg / Flavourzyme® 500 Mg

Flavourzyme 500 Mg

Alcalase 2.4 l Fg / Flavourzyme 500 Mg

A variety of enzymes

can be used to

increase yield and

reduce fermentation

time, as well as to

improve the taste and

aroma of soy sauce.

8

for More iNforMatioNHelen Hu Yingynhu@novozymes.com

The soy sauce made in Vietnam

differs from others because of

the raw materials used. The

special ingredient in Huong Nam Phuong’s

soy sauce is peanut flakes, which are used

instead of wheat.

“Novozymes’ enzymes have helped me

move my production to a healthy level,” says

Thein Cao Ba, owner of Huong Nam Phuong.

Huong Nam Phuong is one of the biggest

soy sauce manufacturers in Vietnam, with an

11% market share. The Vietnamese market is

characterized by many small manufacturers,

most of whom focus on low-cost/high-yield

production. Consequently, many manufac-

turers use acid HVP, leading to the risk of

formation of high levels of 3-MCPD unless

appropriate measures are taken.

“My soy sauce is different from the

others,” says Thein Cao Ba. “Unlike many soy

sauce manufacturers, I follow the Vietnamese

regulations for the amount of 3-MCPD

allowed in soy sauce.”

Only time will tell if Vietnamese consumers

are pleased with Huong Nam Phuong’s soy

sauce made with enzymes. Results so far show

that many customers do appreciate the health-

ier choice. If the trend spreads to the general

population, it will be a tribute to innovative

thinking and sustainable solutions.

vietNaMese soy sauCe a steP ahead of the rest

Huong Nam Phuong, a Vietnamese soy sauce manu-

facturer, is using enzyme technology to produce

healthier soy sauce. Not only is this great for the

company, but consumers gain the best benefit of

all – soy sauce free of dangerous levels of 3-MCPD.

Huong Nam

Phuong uses

both Alcalase®

2.4 l Fg and

Flavourzyme®

500 Mg in its

soy sauce

production.

by one of two methods: acid hydrolysis using

hydrochloric acid or enzymatic hydrolysis using a

balanced mix of exo- and endoproteases. While

acid hydrolysis is cheaper than enzymatic hydroly-

sis, it has major drawbacks.

First of all, 3-MCPD (3-monochloropropane-

1,2-diol) is produced when acid hydrolysis takes

place at high temperatures due to the strong

presence of chloride from the acid. 3-MCPD is a

carcinogen that is banned in food in almost every

country in the world.

Secondly, acid hydrolysis cuts the soy and

wheat into peptides in a random way. This ran-

domness is not optimal for gaining the best flavor

development.

the benefits of enzymesEnzymatic hydrolysis offers the advantage of

eliminating the risk of 3-MCPD formation. This

is because the hydrolysis of the soy and wheat

takes place at closer to neutral pH and at lower

temperatures. This more gentle processing

excludes the formation of any significant level of

3-MCPD.

On top of this, a mix of endo- and exopepti-

dases like Alcalase® 2.4 l Fg and Flavourzyme®

500 Mg provides a blend of peptides, which

gives an excellent flavoring profile. At this point

the hydrolysate can be added to the soy sauce as

a flavoring agent without any further fermenta-

tion. This is the fastest process, since no time is

spent on fermentation afterwards, but even bet-

ter flavors are achieved if the enzymatic hydrolysis

is followed by a fermentation process.

A variety of enzymes can be used at differ-

ent steps in soy sauce production to optimize

both the process and the final product. Despite

the wide variations of ingredients and processes

used, these enzymes can ensure that the soy

sauce sold on supermarket shelves is tasty and

healthy.

A DANgEROUS COMPOUND

3-MCPD (3-monochloropropane-

1,2-diol) is a compound which can

be formed in foods by the reaction of

chloride with lipids at high temperature.

One way it occurs in food products is

in acid-hydrolyzed vegetable proteins

(acid-HVP) when they are produced

using hydrochloric acid at high tem-

peratures. great caution needs to be

taken to avoid formation of 3-MCPD in

foods, because studies show it to be a

carcinogen in laboratory animals.

9

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160

Days

Therapeutic level

Toxicity level

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Therapeutic protein X

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160

Days

Therapeutic level

Toxicity level

Albumin fusion with protein X

Therapeutic protein X

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Courtesy of BioRexis

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In this hypothetical case, the injected biotherapeutic is rapidly cleared from the circulation after administration, which means high dose rates and daily

administration to maintain effective therapeutic levels. By extending the half-life of the biotherapeutic with albufuse® technology, the significantly reduced

dose and reduced frequency of administration give reduced side effects for the same drug because it does not reach the toxicity level in the blood.

days

ther

apeu

tic

ran

ge

therapeutic level

toxicity level

albumin fusion with protein X

therapeutic protein X

Patient benefitsOne advantage for patients is that less frequent

injections are needed. Some biopharmaceuticals

have to be administered by a nurse at home or at

a clinic, so the number of visits can be dramati-

cally reduced by improving the half-life.

Over the course of the treatment, a significant-

ly reduced dose is required, making the treatment

more cost-effective. This is becoming even more

important with the increasing focus on healthcare

costs and accessibility to medicine.

There is also a reduced risk of side effects. A

lower dose rate means that the toxicity level of

the protein may not be reached. Instead, the drug

dose remains within the therapeutic range (see

Fig. 1).

other licenseesAlbuferon is just one of the drugs under develop-

ment featuring albufuse technology. A number

of major biotech and pharmaceutical companies

such as gSk, Teva, CSl Behring, and Dyax are also

using the technology under license.

The latest peptides to awaken great interest are

antibody fragments. These antibody therapeutics

are predicted to represent one of the most excit-

ing and medically beneficial drug development

areas of the next decade. However, development

is being slowed by the high cost of commercial-

scale manufacture using mammalian expression

systems. Novozymes could provide a more cost-

effective solution with a combination of its yeast

expression technology and albufuse technology.

Albumin is the most abun-

dant protein in the human

bloodstream. Novozymes can

fuse pharmaceutical proteins

to albumin to enhance their

half-life. A half-life of hours

becomes days, and days

become weeks.

FIg. 3. A VARIETY OF CONSTRUCTS CAN BE DESIgNED AND MADE

The DNA sequence

for the protein of

choice (shown in

red) can be joined

to albumin in a

number of ways.

C-terminal

N-terminal

In the middle

Combinations

“What a great vehicle!” exclaims Dave Mead,

Business Development Director at Novozymes

Biopharma in Nottingham, Uk. He is not talking

about his favorite car but his favorite carrier –

albumin. “Albumin is present in high volumes in

the bloodstream and it has a naturally long half-life

of 20 days,” he explains. “It doesn’t have an activ-

ity itself apart from being a carrier and that makes

it effectively benign. It’s a great molecule to use as

a carrier for transporting various proteins around

the body.”

These properties make albumin a natural choice

for a drug delivery system. In the mid-1990s,

Novozymes Biopharma succeeded in developing

a proprietary technology known as albufuse® for

genetically fusing target proteins or peptides to

albumin. A typical albumin fusion protein molecule

is shown in Figure 2. The main benefit is that ther-

apeutic proteins become much more effective with

better uptake in the body.

albuferon®* lasts a weekHuman interferon is one example of an albufuse

product. Interferon is administered as a drug to treat

people suffering from a chronic hepatitis C viral

infection. However, with a normal half-life of five

hours, the interferon is soon lost from the body.

One way to make it last longer is to use

PEgylation, whereby a chemical is bolted on to the

interferon. This extends the half-life to 35 hours.

However, by fusing interferon to human albumin,

the half-life becomes 160 hours or one week –

a 32-fold improvement from five hours.

This solution has been chosen by Human

genome Sciences, Inc. in collaboration with

Novartis to make a drug called Albuferon – a com-

bination of albumin and interferon α-2b. Albuferon

is used to treat people with hepatitis C infections.

Human genome Sciences has a license to use the

albufuse technology from Novozymes.

Albuferon requires half the number of injections

compared with PEgylated interferons, and phase 2

results demonstrated that Albuferon offers at least

comparable efficacy, comparable safety, and the

potential for improved health-related quality of life.

Albumin

genetic fusion

FIg. 2. AN AlBUMIN FUSION PROTEIN MOlECUlE OF HUMAN AlBUMIN WITH HUMAN INTERFERON

Interferon α gene

* This registered trademark is the property of Human Genome Sciences, Inc.

EN ©

NO

VO

ZYM

ES A/S D

ATA

gRA

F-2008-19076-01

NovozyMes a/s

krogshoejvej 36

2880 Bagsvaerd

Denmark

Tel. +45 4446 0000

Fax +45 4446 9999

www.biotimes.com

b i o p h a r m a

Antibody–albumin fusions have been shown by

Novozymes to significantly extend the in vivo resi-

dence time of the fragments and they can be made

without the high costs associated with manufactur-

ing monoclonal antibodies.

New possibilities“You can rethink your whole approach and make

completely new therapeutics,” comments Dave

Mead, whose long research background started with

Delta Biotechnology ltd, which was acquired by

Novozymes in 2006. “There are drugs that you could

never produce before without this albufuse scaffold

to support them. And the drugs can be patented,

giving added value to our partners.”

The albufuse concept from Novozymes is suitable

for many peptides and proteins. There are several

binding points on the heart-shaped albumin molecule

where proteins and peptides can be placed.

It is even possible to fuse two different proteins

together with albumin in the same recombinant

molecule to give two different functions at the same

time (see Fig. 3). The new technology is therefore not

just suitable for improving an existing product but

also for making novel products such as bifunctional

proteins.

The researchers at Novozymes have engineered

about 100 different “constructs” and successfully

expressed these fusions.

high yields with yeastThe fusion technique is based on fusing the DNA

encoding for short-lived peptides or proteins to the

DNA encoding for albumin. The recombinant DNA

is engineered into the host cell Saccharomyces

cerevisiae, known more commonly as baker’s yeast,

which has been used for centuries in breadmaking,

winemaking, and brewing. This microorganism is

well understood and is generally recognized as safe

(gRAS). The fusions are secreted with high yields in

a fermentation tank and then purified to a high level

of purity.

Another advantage of albumin fusion proteins is

that they are produced in a system free of animal

components.

license packagesNovozymes has 10 years’ experience of producing

recombinant human albumin (rHA) using its proprie-

tary yeast expression technology at its cgMP (current

good Manufacturing Practice) manufacturing plant

in Nottingham in the Uk. The company’s experts can

help with license packages to develop drugs pro-

duced using this expression technology in combina-

tion with the albufuse technology.

“We’ve made many types of proteins over the

years and they have all done remarkably well. We

have a number of licenses already and we’ve per-

formed technology transfer with a number of third

parties across the globe,” says Dave Mead. “Our

yeast expression system has been proven to produce

pharmaceutical-grade products.

“At Novozymes, we can take our Biopharma cus-

tomers a long way up the learning curve. We can

give them access to our tremendous expertise, our

patents, and our extensive series of yeast strains. We

also offer contract manufacturing of pharmaceutical-

grade biological drugs at our cgMP facility at lund

in Sweden. We are not a drug discovery company

but we work with partners from initial protein

expression through process development work to

delivery of a commercial-scale bioprocess that is

robust, reproducible, and cost-effective. This will help

them get to the marketplace faster, which is where

the pot of gold lies at the end of the day,”concludes

Dave Mead.

for More iNforMatioNDave Meaddvjm@novozymes.com

“The type of peptide or pro-

tein you choose and where

you place it on the albumin

molecule depend on what you

want to do. It’s a powerful

technology,” says Dave Mead,

Business Development Director

at Novozymes Biopharma.

CONgRATUlATIONS TO

Agustina Mezquita, Michael Porter, and

Ming Huang, the three winners of the

iSongBook. Thanks to all of you who

participated in our BioTimes survey.