PRODUCINGCEPHALOSPORIN

In 1929,Alexander Fleming had discovered Peniciline

SETTING PROBLEM

CONTENTSIII. Producing Cephalosporin C

IV. Application.

II. About C

ephalosporin.

I. G

ener

al v

iew a

bout A

ntibio

tics.

PART I: GENERAL VIEW ABOUT ANTIBIOTICS.

To inhibit the growth of or to destroy bacteria

Have a specific

Low concentration

Secreted by microoganism

synthetic or semi-synthetic chemical compound

AntibioticsAnti

biotics

I.DEFINITION

The antibiotics have β-lactam structure Penicillin,cephalosporin

The antibiotic containing aromatic ring Chloramphenicol

Aminoglycosid structure antibiotics Streptomycin,gentamicin

The structures of antibiotics have 4 rings Tetracyclins

Polypeptid antibiotics Polymyxin,bacitracin

Macrolid antibiotics Erythromycin,spiramycin

Polyen antibiotics Nystatin,amphotericin B

………………..

II.CLASSIFICATION TABLE

PENICILINES

CEPHALOSPORIN

MONOBACTAMS

CARBAPENEMS

III.β- LACTAM GROUP

-lactams Group

-lactamsrings

Antibiotic resistance -lactams group

RESIST BY β-LACTAMASE ENZYME

PART II: GENERAL VIEW ABOUT

CEPHALOSPORIN ANTIBIOTICS.

1945

1956

1964

1971DISCOVERED HISTORY

Cephalosporium acremonium race was discovered from a sewer in Sardinia in 1945 by Italian scientist Giuseppe Brotzu

Cephalosporin C and penicillin N were fractionated from fungus

growth sap

First introduced in clinal in 1964 (cephalothin)

Cephamycin was fractionated fromNorcadia race, having same structure as Cephalosporin

Repressing cell wall biosynthesis

One of β-lactam antibiotics

Operating on Gr(+) and Gr(-) bacteria

CEPHALOSPORIN

1Nature CP have low antibacterial active

2Cephalosporin includes β-lactam ring and dihydrothiazin heterocyclic

(cephem core)

3Cephalosporin use for treatment all are synthesis or semi-synthesis chemical compound

4Cephalosporin is the material to

produce 7-ACA and 7-ADCA

CEPHALOSPORIN ’S GENERAL

CEPHALOSPORIN’S GENERATIONS

1st gener.

Sensitive with β-lactamase.

+Cefazolin+Cephalexin+Cepoxitin

2nd gener.Sensitive with

β-lactamase.

+Cefamadole+Cefaclor+Cepoxitin

3rd gener.

β-lactamase inhibit.

+Cefotaxime+Ceftriaxone

4th gener.

β-lactamaseinhibit, Repress Pseudomoas

+Cefepime+Cefozopran

5th gener.

β-lactamaseinhibit,Repress MRSA

+Ceftobiprole+Ceftaroline

Cep

hal

osp

orin

cl

assi

fica

tion

Name

PART III: PRODUCING CEPHALOSPORIN C BY

MICRORGANIC FERMENTATION METHOD

I.CEPHALOSPORIN C (CPC) BIOSYNTHESIS

1. Race. - Cephalosporium acremonium (Acremonium chrysogenum)

- Low antibiotics active, doesn’t have super-synthetic race.

- Doesn’t have acyltransferase enzyme so can’t use precursor substance to contribute new product

Pictures of Cephalosprium Acremonium race

Cephalosporin acremonium

Ingredient:

1.3. Growth condition:

Nutrition

Microquantity

pH

Water

Sacharose 36gGlucose 27g(NH4)2SO4 7.5gOleic acid 1.5gMethionin 3.0g

K, Fe, Mn, Mg, Zn, Cu,…

7.3

(Adequate add 1liter)

1.4 Fermentation condition:

pH condition from 6,8 to 7,4.

Temp. pH Oxigen CO2

.

T = 25-300C, maintain during fermentation process

Concentration of dissolved oxygen = 30% concentration of saturated one.

Too high concentration of CO2 will resist the adsorbtion and metabolization of substrate ‘s races.

1.5 Prepare for fermentation:

Conserve races and multiplicate races: Extremely cold storage at -700C or liquid nitrogen

shielding. Races from preservation condition are

transplanting to agar to active and growth to get spore.

Finally, the multiplicating equipment will produce the amount of races whose the density in initial ferment solution is about 1 - 5.109 spores / m3.

•Ferment equipment: Must be in absolute sterility before using, either to all of the equipments. Sterile in overheated steam at 2,5 – 3,0 at for 3 hr. Manage to maitain the excess pressure in the equipment during the ferment process to limit contaminant.

•The air is usually been antiseptic by adiabatic compresion, then get through ultrafiltration.

•Surfaced fermentation engineering: Fermenting on solid materials (corn mash, wheat mash added lactose)

BIOSYNTHESIS MECHANISM

4. Fractionate, purify

6.Mass produce

1.Subdividing,Selecting race

2.Preserving, activating race

3. Produced ferment

PROCESS

3.Producing Process:

5. Collecting product

Step 1. Subdeviding and selecting race

5.Transplant bacteria into Mueller-Hinton agar disk

6. Put in antibiotic paper

7. Measure absolute sterility ring

8.Analyse and answer result

1.Subdevidingrace

2. Transplant into environment without selection

3. Creating bacteria sap in normal saline (108cell/ml)

4.Adjusting opacity of bacteria (106cell/ml)

Cephalosporin acremonium

UI

Step 3: Produced Ferment

YBS environment ( growth environment) Two phases fermentFirst phase (get biomass): 2-3 days, at 28 – 30oC, pH 6.5-6.8Last phase (get product) : 5 -7 days, at 22-25oC, pH lower than frist phase

Step 4: Frationate and Purify

Fermentation method:

Ferment in batch-reactor

Using stirrer blades

Aerating air by power pump

Producing CEPHALOSPORIN fermentation figure

Submerged fermentation

Foam breaking

Partition

Heat transfer

Flat turbine

Aseptic air

EQUIPMENT FACTOR

Volume : 5 m3 Filled coefficient : 0.75 Steel component: X18H10T and X17H13M2T Stir speed: 110-200 rpm Antiseptic at: 130- 140ºC Working pressure : 50 kPa – 0.25 Mpa Working temprature: 0 – 34ºC Blow rate : 12 m/s

FERMENT PROCESS

Ferment equipment inside laboratory

Pilot

Industrial Fermentation

PLEASE KEEP TRACK OF THIS FOLLOWING VIDEO

Visit website: http://www.biocon.com/bioconvirtualtour_v1/submergedfermentation.htm

5. CALCULATING KINETIC FACTORS OF EQUIPMENT:

Bacterial growth curve in the batch reactor

Reaction scheme

= k . = max.

CC Bacterial concentration (biomass)(g/l) µ particular day growth speed (day-1) µmax maximun particular day growth speed(day-1) CA sucrose concentration (g/l) CM

Monod coefficient (g/l).

Monod equation for batch reactor:

rC = = µCC =

Calculating data

CA (g/l) 3 2.9 1.7 1.2 0.8 0.3 0.2

CC (g/l) 0 0 0.2 0.33 1 1.4 1.6

t 0 1 2 3 4 5 6

0 1 2 3 4 5 60

1

2

3

4

5

6

7

8

9

f(x) = 1.41996987793073 x + 0.142586087819977R² = 0.921047635800439

1/CA 0.59 0.83 1.25 3.3 5

Cc/rc 1 1.57 2.22 3.7 8

Reaction rate

We have this from the chart above:K= 7 day-1

Cm= 9.94 (g/l)

Volume of equipment

Antibiotics concentration: 1.6 g/l.Weekly yeild: 30 kgAnually yeild: 1560 kgVolume of equipment 18.75 m3

Need 5 serial reactors

With this ouput, about 150.000600.000 Cefpodoxime tablet will be produced per week.

Part IV: Appication

Nowadays, Cephalosporin is the most important antibiotic drug. Cephalosporin use to treat a lot of disease, what resisted another antibiotics such as: penicilin, amoxicilin, ampicilin…

Ranking 7th / 10 antibiotics to treat infections, it is caused by staphylococus aureus and typhoid fever in humans.

Semi-synthetic CPC have antibacterial effect with both β-lactamase bacteria, gram (-) bacteria and have low toxicity.

›CPC second generation antibiotics is very specially effect when we use infections of incision after operation.

›Advantage of CPC third generation antibiotics is the most broad spectrum antibacterial for both bacteria gram (-) and gram (+),including strains of streptococcus .

›It used to treat severely infections in bronchus and lung, sepsis, bacterial contamination of bone, urinaly tract, gonorrhoeae resisted by another antibiotics…

1. Nguyễn Thị Mai 609015252. Phạm Thị Hồng Nhật 609018443. Thiều Nguyễn Trường Giang 609006804. Vũ Viết Văn Thưởng 608021995. Phạm Như Quốc Khánh 609011926. Hứa Tiến Đức 609006257. Trần Ngọc Hưng 609011198. Nguyễn Văn Nguyên 609017649. Trần Ngọc Kiều Khanh 60901164

MEMBERS

INSTRUCTOR :Doctor. Lê Thị Kim Phụng

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