Pilot scale biotransformation of vegetal oil into natural green note flavor using sugar beet leaves as sources of hydroperoxide lyase

(1)

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

leaves as sources of hydroperoxide lyase

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

leaves as sources of hydroperoxide lyase

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Introduction

_{Plant materials characterization}

_{Aldehyde synthesis}

_{Proposed pilot-scale biotransformation process of vegetal}

Introduction

Natural green note aromas (GLVs) are highly attractive flavors

Plant materials characterization

_{Nowadays sugar beet leaves are a unvalued by-product of the}

Aldehyde synthesis

Proposed pilot-scale biotransformation process of vegetal

Natural green note aromas (GLVs) are highly attractive flavors

commonly used in the food industry. These are produced in Nowadays sugar beet leaves are a unvalued by-product of the

Aldehyde synthesis

This is the most critical step of the lipoxygenase pathway which

Proposed pilot-scale biotransformation process of vegetal

oil into green note flavor using sugar beet leaves

commonly used in the food industry. These are produced in Nowadays sugar beet leaves are a unvalued by-product of the_{agricultural industry in Europe. This raw plant material} This is the most critical step of the lipoxygenase pathway which

is due to the instability of hydroperoxide lyase activity. The

oil into green note flavor using sugar beet leaves

commonly used in the food industry. These are produced in

extremely low levels upon physiological stress in plant organs agricultural industry in Europe. This raw plant material_{presents valuable enzymatic content, like hydroperoxide lyase} is due to the instability of hydroperoxide lyase activity. The

aldehyde synthesis has to be carefully controlled. After the setup

oil into green note flavor using sugar beet leaves

extremely low levels upon physiological stress in plant organs

of any sort. This weak sporadic presence entails a very presents valuable enzymatic content, like hydroperoxide lyase_{activity. Specific characterization of the potential GLVs} aldehyde synthesis has to be carefully controlled. After the setup

of any sort. This weak sporadic presence entails a very

expensive extraction step to obtain pure GLVs. Therefore activity. Specific characterization of the potential GLVs

aldehyde synthesis has to be carefully controlled. After the setup of the optimum conditions (pH and temperature) for the

expensive extraction step to obtain pure GLVs. Therefore activity. Specific characterization of the potential GLVs_{production for these enzymes has led to the following result.} of the optimum conditions (pH and temperature) for the

hydroperoxide lyase of sugar beet, ground plant materials are expensive extraction step to obtain pure GLVs. Therefore

catalytic biotransformations of fatty acid sources, the initial production for these enzymes has led to the following result. hydroperoxide lyase of sugar beet, ground plant materials are_{quickly added to the production juice containing previously}

catalytic biotransformations of fatty acid sources, the initial

substrate for GLVs, have been developed. Enzymatic defense 7,00 quickly added to the production juice containing previously

substrate for GLVs, have been developed. Enzymatic defense 7,00 ₁₄

O x y li p in l e v e l (n m o l/ g o f fr e s h g o

f quickly added to the production juice containing previously

synthesized hydroperoxides. substrate for GLVs, have been developed. Enzymatic defense

pathways and particularly the LOX pathway produce the

14 6,00 O x y li p in l e v e l (n m o l/ g o f fr e s h U E * / g o f synthesized hydroperoxides. pathways and particularly the LOX pathway produce the

major part of GLVs. Unlike GLV molecules that are emitted in 12

6,00 O x y li p in l e v e l (n m o l/ g o f fr e s h U E * / 100%

major part of GLVs. Unlike GLV molecules that are emitted in

the atmosphere, the enzymes are extractible from the plant 5,00

O x y li p in l e v e l (n m o l/ g o f fr e s h a c ti v ty (U E * / 90% 100% P ro d u c ti o n l e v e l (% )

the atmosphere, the enzymes are extractible from the plant 10

5,00 O x y li p in l e v e l (n m o l/ g o f fr e s h a c ti v ty w e ig h t) _80%90% P ro d u c ti o n l e v e l (% )

the atmosphere, the enzymes are extractible from the plant

material. Thus, a combination of plant enzyme extracts and 4,00 ₈

O x y li p in l e v e l (n m o l/ g o f fr e s h w e ig h t) a c ti v ty w e ig h t 70% 80% P ro d u c ti o n l e v e l (% )

material. Thus, a combination of plant enzyme extracts and substrate preparations provides all the ingredients for GLV

8 4,00 O x y li p in l e v e l (n m o l/ g o f fr e s h w e ig h t) H y d ro p e ro x id e l y a s e a c ti v ty w e ig h t 60% 70% P ro d u c ti o n l e v e l (% )

substrate preparations provides all the ingredients for GLV

production. Besides, sugar beet leaves present high levels of 6

3,00 O x y li p in l e v e l (n m o l/ g o f fr e s h w e ig h t) H y d ro p e ro x id e l y a s e fr e s h w e ig h t 50% 60% P ro d u c ti o n l e v e l (% )

production. Besides, sugar beet leaves present high levels of 6

3,00 O x y li p in l e v e l (n m o l/ g o f fr e s h w e ig h t) H y d ro p e ro x id e l y a s e fr e s h 40% 50% P ro d u c ti o n l e v e l (% )

production. Besides, sugar beet leaves present high levels of

hydroperoxide lyase among plant sources and are available in 2,00 4

O x y li p in l e v e l (n m o l/ g o f fr e s h H y d ro p e ro x id e l y a s e fr e s h 30% 40% P ro d u c ti o n l e v e l (% )

hydroperoxide lyase among plant sources and are available in large amounts during three months. In this enzymatic

4 O x y li p in l e v e l (n m o l/ g o f fr e s h H y d ro p e ro x id e l y a s e 20% 30% P ro d u c ti o n l e v e l (% )

large amounts during three months. In this enzymatic pathway, fatty acids are successively transformed by lipase,

2 1,00 O x y li p in l e v e l (n m o l/ g o f fr e s h H y d ro p e ro x id e l y a s e 0% 10% 20% P ro d u c ti o n l e v e l (% )

pathway, fatty acids are successively transformed by lipase,

2 O x y li p in l e v e l (n m o l/ g o f fr e s h H y d ro p e ro x id e l y a s e 0% P ro d u c ti o n l e v e l (% )

pathway, fatty acids are successively transformed by lipase,

lipoxygenase and hydroperoxide lyase into aldehydes and 0,00 0

H y d ro p e ro x id e l y a s e 0 5 10

lipoxygenase and hydroperoxide lyase into aldehydes and

alcohols, final compounds of GLVs pathway. Limiting and 9HPOD 9HPOT 13HPOD 13HPOT

0 5 10

Time (minutes)

alcohols, final compounds of GLVs pathway. Limiting and 9HPOD 9HPOT 13HPOD 13HPOT

Hydroperoxide isomer substrate Time (minutes)

alcohols, final compounds of GLVs pathway. Limiting and

problematic steps occur with the action of hydroperoxide Hydroperoxide isomer substrate _{Fig. 3 : Evolution of the global aldehyde level within the first ten minutes after}

problematic steps occur with the action of hydroperoxide

lyase, when enzymatic catalysis is followed by an enzyme Fig. 1 : Degrading hydroperoxide activity in sugar beat leaves. Bars

Fig. 3 : Evolution of the global aldehyde level within the first ten minutes after the addition of sugar beet leaves

lyase, when enzymatic catalysis is followed by an enzyme destabilization. Alternative substrates bind irreversibly to the

Fig. 1 : Degrading hydroperoxide activity in sugar beat leaves. Bars represent the hydroperoxide lyase activity related to the four hydroperoxide

the addition of sugar beet leaves

destabilization. Alternative substrates bind irreversibly to the represent the hydroperoxide lyase activity related to the four hydroperoxide

substrates. Lines show the concentration of corresponding hydroperoxide _{Following the mixing} _{of substrates and enzymes, only}

destabilization. Alternative substrates bind irreversibly to the

heme group of the enzyme and end the reaction. This poster substrates. Lines show the concentration of corresponding hydroperoxide_{isomer in the leaves.} Following the mixing of substrates and enzymes, only

productions of Z-3-Hexenal and E-2-Hexenal were monitored. heme group of the enzyme and end the reaction. This poster

briefly describes the development of a complete bioprocess

isomer in the leaves. Following the mixing of substrates and enzymes, only

productions of Z-3-Hexenal and E-2-Hexenal were monitored. briefly describes the development of a complete bioprocess

for natural GLV production, from hydrolysis to purification. A The hydroperoxide substrates naturally present in leaves are

productions of Z-3-Hexenal and E-2-Hexenal were monitored. Maximum reaction levels are obtained within 5 minutes and

for natural GLV production, from hydrolysis to purification. A The hydroperoxide substrates naturally present in leaves are

clearly linolenic acid derivate : 9-HPOT and 13-HPOT.

Maximum reaction levels are obtained within 5 minutes and thereafter decrease slowly due to the aldehyde consumption by for natural GLV production, from hydrolysis to purification. A

high level of biotransformation could be achieved using clearly linolenic acid derivate : 9-HPOT and 13-HPOT.

Although concentrations are very low, this indicates that

thereafter decrease slowly due to the aldehyde consumption by high level of biotransformation could be achieved using

optimum experimental conditions and a cheap source of plant Although concentrations are very low, this indicates that

thereafter decrease slowly due to the aldehyde consumption by other enzymatic activities from the plant materials. The optimum experimental conditions and a cheap source of plant

materials.

Although concentrations are very low, this indicates that sugar beet have preferentially developed this way of

other enzymatic activities from the plant materials. The maximum concentration can be maintained by pH reduction to

materials. sugar beet have preferentially developed this way of

synthesis. Furthermore, only 13-hydroperoxide lyase activity

maximum concentration can be maintained by pH reduction to inhibit the enzymatic degradation.

materials.

synthesis. Furthermore, only 13-hydroperoxide lyase activity _{inhibit the enzymatic degradation.}

synthesis. Furthermore, only 13-hydroperoxide lyase activity has been found. Thus, Z-3-Hexenal and E-2-Hexenal,

inhibit the enzymatic degradation. has been found. Thus, Z-3-Hexenal and E-2-Hexenal,

resulting from the activity of 13-hydroperoxide lyase on 13- ₎ 0,25 90,00%

resulting from the activity of hydroperoxide lyase on

13-HPOT, are the preferential compounds we are going to 80,00%

90,00% 0,25 B io tr a n s fo rm a ti o n Y ie ld . 1 0 m in )

Objective and purpose

HPOT, are the preferential compounds we are going to

70,00% 80,00% 0,2 B io tr a n s fo rm a ti o n Y ie ld . 1 0 m in

Objective and purpose

HPOT, are the preferential compounds we are going to

investigate for flavor production with sugar beet leaves. 60,00%

70,00% 0,2 B io tr a n s fo rm a ti o n Y ie ld / L . 1 0 m in

Objective and purpose

Establishing a simple production bioprocess of green leaf

investigate for flavor production with sugar beet leaves. 60,00%

0,15 B io tr a n s fo rm a ti o n Y ie ld m m o l/ L

Establishing a simple production bioprocess of green leaf flavors using unwanted plant materials like sugar beet leaves.

50,00% 0,15 B io tr a n s fo rm a ti o n Y ie ld (m m o l

flavors using unwanted plant materials like sugar beet leaves. 40,00%

0,1 B io tr a n s fo rm a ti o n Y ie ld le v e ls

flavors using unwanted plant materials like sugar beet leaves.

The process has to be efficient and leads to the purest flavor 30,00%

0,1 B io tr a n s fo rm a ti o n Y ie ld P ro d u c ti v it y le v e ls

The process has to be efficient and leads to the purest flavor

composition.

Fatty acids hydrolysis

0,05 20,00%

B io tr a n s fo rm a ti o n Y ie ld P ro d u c ti v it y

composition.

Fatty acids hydrolysis

0,05 _10,00%20,00%

B io tr a n s fo rm a ti o n Y ie ld P ro d u c ti v it y

composition.

Fatty acids hydrolysis

Following plant characterization, linseed oil has been chosen 0,00%

10,00% 0 B io tr a n s fo rm a ti o n Y ie ld P ro d u c ti v it y

Following plant characterization, linseed oil has been chosen as substrate source, on account of its high linolenic acid

0,00% 0 0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2 P ro d u c ti v it y

as substrate source, on account of its high linolenic acid content. Hydrolysis is a common chemical reaction in industry.

0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2

Substrate concentration (mM)

content. Hydrolysis is a common chemical reaction in industry. Substrate concentration (mM)

Material and Methods

content. Hydrolysis is a common chemical reaction in industry.

We hydrolyze the oil enzymatically with lipase to keep neutral _{Fig. 4 : Evolution of productivity (grey) and yield (brown) levels of Z-3-Hexenal}

Material and Methods

All reactions have been performed in pilot scale vessels,

We hydrolyze the oil enzymatically with lipase to keep neutral condition at room temperature. Hydrolysis is performed by

Fig. 4 : Evolution of productivity (grey) and yield (brown) levels of Z-3-Hexenal related to 13-hydroperoxide substrate concentration.

All reactions have been performed in pilot scale vessels, condition at room temperature. Hydrolysis is performed by related to 13-hydroperoxide substrate concentration.

All reactions have been performed in pilot scale vessels, including mixed bioreactor of 100L. Reaction order simply

condition at room temperature. Hydrolysis is performed by immobilized lipases from a Bacillus strain. Enzymes are including mixed bioreactor of 100L. Reaction order simply

follows the natural lipoxygenase pathway present in most of

immobilized lipases from a Bacillus strain. Enzymes are

recycled 3 times and a high yield (more than 90%) of free High GLVs concentrations are difficult to obtain because of the

follows the natural lipoxygenase pathway present in most of recycled 3 times and a high yield (more than 90%) of free

fatty acids has been recovered after centrifugation. No

High GLVs concentrations are difficult to obtain because of the

inhibition of hydroperoxide lyase above 0,15 mM of

follows the natural lipoxygenase pathway present in most of

the superior plants. fatty acids has been recovered after centrifugation. No inhibition of hydroperoxide lyase above 0,15 mM of

hydroperoxide substrate. But further experiments with substrate

the superior plants. fatty acids has been recovered after centrifugation. No

emulsifying compounds have been used to avoid loss of free hydroperoxide substrate. But further experiments with substrate

emulsifying compounds have been used to avoid loss of free fatty acids by centrifugation.

hydroperoxide substrate. But further experiments with substrate concentrations above 2mM have shown better productivity levels

fatty acids by centrifugation. concentrations above 2mM have shown better productivity levels

in spite of a lower yield. in spite of a lower yield.

Main substrate source

Fatty acids oxidation

Main substrate source

Sugar beet leaves do not present high levels of fatty acids

Fatty acids oxidation

Sugar beet leaves do not present high levels of fatty acids content (only 0,1%). Therefore, the use of an external source

Fatty acids oxidation

The lack of LOX activities (less than 0,1 UE*/ g of fresh

_{Downstream Processing}

content (only 0,1%). Therefore, the use of an external source of initial substrate is necessary. Vegetal oils are cheap sources

_{Downstream Processing}

of initial substrate is necessary. Vegetal oils are cheap sources

weight) in sugar beet leaves requires an external source of this

Downstream Processing

After the synthesis steps, an optional alcoholic conversion of of initial substrate is necessary. Vegetal oils are cheap sources

of unsaturated fatty acids which are easily obtained, and

weight) in sugar beet leaves requires an external source of this

enzyme. Actually, crude soybean seeds are the best sources of After the synthesis steps, an optional alcoholic conversion of

of unsaturated fatty acids which are easily obtained, and stable enough for flavor production. Final aroma composition

enzyme. Actually, crude soybean seeds are the best sources of 13-lipoxygenase activity. This reaction is fast and completed

After the synthesis steps, an optional alcoholic conversion of aldehydes into alcohols can be performed, but in our case no

stable enough for flavor production. Final aroma composition 13-lipoxygenase activity. This reaction is fast and completed aldehydes into alcohols can be performed, but in our case no

transformation was made. Firstly, the production juice is cleared stable enough for flavor production. Final aroma composition

will depend on concentration of linoleic or linolenic acid

13-lipoxygenase activity. This reaction is fast and completed

within 30 minutes at 4°C and pH 9.3. transformation was made. Firstly, the production juice is cleared

of plant fragments, by filtration through 0.5mm filters upon 1 bar will depend on concentration of linoleic or linolenic acid

present in the oil.

within 30 minutes at 4°C and pH 9.3.

of plant fragments, by filtration through 0.5mm filters upon 1 bar present in the oil.

100 70

of plant fragments, by filtration through 0.5mm filters upon 1 bar of pressure. To recover aldehydes from the juice, adsorption on

90 100 60 70 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o

n of pressure. To recover aldehydes from the juice, adsorption on

ionic resin has been adapted. The MN-202 resin from purolite,

80 90 60 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n

ionic resin has been adapted. The MN-202 resin from purolite, usually used for water cleansing, presents perfect characteristics

usually used for water cleansing, presents perfect characteristics

usually used for water cleansing, presents perfect characteristics and cheap prices to accomplish the adsorption. 95% of the

50 30 40 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n (m M

) and cheap prices to accomplish the adsorption. 95% of the

aldehydes are effectively adsorbed and after that 75% are eluted

Conclusion

40 30 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n (m M )

aldehydes are effectively adsorbed and after that 75% are eluted

in isopropanol or a different alcoholic solvent. Secondly, steam

Conclusion

30 20 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n (m M )

Conclusion

At the end of the process, pure aldehydes (Hexanal and E-2-Hexenal) can be recovered. These

Tab 1. Synthesis potential for different vegetal oils related to their maximum

20 10 O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n

distillation is used to purify the flavor juice by evaporating At the end of the process, pure aldehydes (Hexanal and E-2-Hexenal) can be recovered. These

Tab 1. Synthesis potential for different vegetal oils related to their maximum 10 10

O x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n

distillation is used to purify the flavor juice by evaporating alcohol. No azeotropic gas was observed during distillation and

At the end of the process, pure aldehydes (Hexanal and E-2-Hexenal) can be recovered. These compounds can be labeled as “natural flavor” because they are vegetal oil derivates. The price value is

level of C₆Aldehydes production (mg/g of oil) 0 0 O

x y g e n s a tu ra ti o n ( % ) H y d ro p e ro xi d e c o n c e n tr a ti o n

alcohol. No azeotropic gas was observed during distillation and all aldehydes could be conserved. But, our apparatus was not

compounds can be labeled as “natural flavor” because they are vegetal oil derivates. The price value is more than 1000$/kg for these compounds and they could be easily sold in food and beverage sectors.

6 0 5 10 15 20 25 30 H y d ro p e ro xi d e c o n c e n tr a ti o n

all aldehydes could be conserved. But, our apparatus was not more than 1000$/kg for these compounds and they could be easily sold in food and beverage sectors.

This long and complex biotransformation of cheap resources into expensive flavors may be a solution to

These oils are highly valorized through this bioprocess because Time of reaction (minutes)

all aldehydes could be conserved. But, our apparatus was not

able to separate hexanal from hexenal isomers. Also, during This long and complex biotransformation of cheap resources into expensive flavors may be a solution to

These oils are highly valorized through this bioprocess because this aroma is sold at more than one hundred times the price of

Time of reaction (minutes) _{able to separate hexanal from hexenal isomers. Also, during}

distillation it was observed that all Z-3-Hexenal isomers were

This long and complex biotransformation of cheap resources into expensive flavors may be a solution to convert unwanted crop byproducts, such as sugar beet leaves.

this aroma is sold at more than one hundred times the price of _Fig. ₁ _: _{Biotransformation} _of _100mM _linolenic _acids _solution _into distillation it was observed that all Z-3-Hexenal isomers were convert unwanted crop byproducts, such as sugar beet leaves.

this aroma is sold at more than one hundred times the price of

the oil. But only C₁₈ unsaturated acids of the oil are Fig.hydroperoxide by 13-Lipoxygenase. Grey line represents the hydroperoxide1 : Biotransformation of 100mM linolenic acids solution into

distillation it was observed that all Z-3-Hexenal isomers were totally converted into E-2-Hexanal.

the oil. But only C₁₈ unsaturated acids of the oil are

metabolized through this enzymatic pathway.

hydroperoxide by 13-Lipoxygenase. Grey line represents the hydroperoxide concentration during reaction, and brown line the oxygen level in bioreactor.

totally converted into E-2-Hexanal.

metabolized through this enzymatic pathway. concentration during reaction, and brown line the oxygen level in bioreactor.

Pilot scale biotransformation of vegetal oil into natural green note flavor using sugar beet leaves as sources of hydroperoxide lyase

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

leaves as sources of hydroperoxide lyase

Pilot scale biotransformation of vegetal oil into natural green notes flavor using sugar beet

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

leaves as sources of hydroperoxide lyase

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Gigot C., Ongena M., Fauconnier M.-L., Wathelet J.-P., Du Jardin P. And Thonart P.

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Walloon Center of Industrial Biology, Gembloux argicultural University, Passage des déportés 2, 5030 Gembloux, Belgium

Introduction

Introduction

Plant materials characterization

Aldehyde synthesis

Proposed pilot-scale biotransformation process of vegetal

Introduction

Plant materials characterization

Aldehyde synthesis

Proposed pilot-scale biotransformation process of vegetal

Aldehyde synthesis

Proposed pilot-scale biotransformation process of vegetal

oil into green note flavor using sugar beet leaves

oil into green note flavor using sugar beet leaves

oil into green note flavor using sugar beet leaves

Objective and purpose

Objective and purpose

Objective and purpose

Fatty acids hydrolysis

Fatty acids hydrolysis

Fatty acids hydrolysis

Material and Methods

Material and Methods

Main substrate source

Main substrate source

Fatty acids oxidation

Main substrate source

Fatty acids oxidation

Fatty acids oxidation

Downstream Processing

Downstream Processing

Downstream Processing

Conclusion

Conclusion

Conclusion

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

* 1UE corresponding to 1mM of hydroperoxide converted by minutes

_{Plant materials characterization}

_{Aldehyde synthesis}

_{Proposed pilot-scale biotransformation process of vegetal}

_{Downstream Processing}

_{Downstream Processing}