Assalammualaikum and avery good evening.

I will present my project entitled, “xylitol production of recombinant E.coli

immobilized on MWCNT”

Firstly, what is xylitol??

1) Xylitol is a sugar alcohol that mainly used as sweetener and benefits to

diabetic people.
2) Xylitol can be produced from xylose by 2 different method which are by
chemical approach and biological approach.
3) Xylose is extracted from biomass by chemical pretreatment and hydrolysis.
4) In my study, I focused the xylitol production by bacteria.

1) Now, this is the previous studies for the xylitol production by several
microorganism as listed here.
2) The highest xylitol productivity was obtained by using Candida tropicalis.
3) Some literature claimed that, Candida tropicalis is the best producer for
xylitol.

1) In this project, Recombinant E. coli was chosen as a host for xylitol

production.
2) This is because E. coli has some advantages such as:
- Is an efficient biocatalyst
- Well established genetic manipulation

3) The enzyme that responsible for xylitol production in E.coli is XR.

4) Naturally, E. coli unable to produce xylitol.
5) Thus, XR gene from neurospora crassa was cloned into E.coli to enable
xylitol production.
6) In E.coli, xylose is metabolized into energy by 2 enzyme xylA (xylose
isomerase) in Pentose Phosphote Pathway.
7) Therefore, the deletion of xylA and xylB enzyme was done in order to direct
xylose metabolism only for xylitol production by using XR enzyme.
1) There are some problems in xylitol production especially using free cells.
2) When used bacteria E.coli, the common problem faced are low yield, high
cell lysis and low plasmid stability.
3) The use of cell immobilization technique is expected to improve xylitol
production, plasmid stability and reduce cell lysis.

1) The basic concept of cell immobilization is the localization of cell to a

certain region of immobilization matrix with the desired catalytic activity.

1) The cell immobilization technique can be divided into 2, first: entrapment,

second: adsorption
2) Entrapment method such as matrix encapsulation and microencapsulation.
For this method, cell usually embedded into the matrix and not directly
contact with nutrients.
3) Meanwhile, for adsorption method such as physical adsorption onto surface
of immobilization matrix and chemical adsorption via covalent bond.
4) And in this study, I used natural adsorption technique.
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1) I chose adsorption method because of some advantages as stated in these

previous literatures such as low cost and allows an efficient binding process.
However, significant leakage rate is one of the drawback for the technique.

1) This is the cells immobilized on MWCNT by adsorption .

2) This is the previous studies that use CNT as immobilization matrix for many
purposes. By using CNT, it could provide some advantages such as large
surface area, increased cell stability and high reusability.
1) My research objective is To improve the xylitol production, cell viability
and plasmid stability by immobilizing E. coli on multiwalled carbon
nanotubes (MWCNT) using optimum cultural conditions.
2) My research scope divided into 3:
a. Screening effect the MWCNT treatment on cell immobilization
b. Screening effect of cultural conditions on xylitol production using
OFAT.
c. Screening effect of cultural condition on xylitol production using
RSM