MPH TAXUS_BACCATA

8/6/2019 MPH TAXUS_BACCATA

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2. Cultivation ofTaxus baccata,

Ginseng, Artemisia annua, (08)


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CULTIVATION AND COLLECTION OF

TAXUSBACCATA


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PLANTPROFILE:-

Synonym:- Yew, Talispatra.

Biological Source:- Taxol is a naturally occuring diterpenoid belon

ging to taxane group of compounds present

in genus Taxus.

Family:- Taxaceae.

Various Species:- Taxus baccata (English or European)

Taxus cuspidata( japanese)

Taxus brevifolia(Pacific Yew)Taxus canadensis(Ground hemlock,

canadian,American yew)

Chemical constituents:- Taxanes are important group of chemical

constituents ,cephalomannine,10- deacetyl baccatin.


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Uses:- Refractory ovarian cancer

Non-small cell lung carcinoma

Gastric and Cervical cancers

Carcinomas of head,neck,prostate and colon.

The derivative of taxol called taxotere has better bioavaili

bility and has been claimed as a promising anticancer

agent.


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CULTIVATION OF TAXOL:-


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SOIL & CLIMATE:-

Sandy soil, light brown lime soil,dry soil but flourish inwet loam drained soil slightly alkaline.

It needs moist temperate climate.

LAND PREPARATION:-

Field should be prepare to make good seed beds,bylight ploughing then followed by breaking the clumpson the earth before the onset on the monsoonseasons.


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MODESOFPROPAGATION:-

Propagated by seeds and shoots cutting.

NURSERYRAISING:-

The best time for nursery raising is May- June.

Seeds require shelter and moist shady area for germination and donot survive in open areas.

The seedlings get prepared in early april when sprouts comes outfrom the surface the beds are covered and protected from sunlight.

The growth continues from the stage of germination throughoutsummer.


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In the September or early October the leaves and the stem wiltoff and show the covering of the stock with soil is necessary inorder to develop the underground stem and to protect root from

the frost in the late may or in early june next year the seedlingsare dug out and transplanted in the field.

TRANSPLANTING:-

Next year before onset of monsoon the taxus seedling areplanted in the field.

The trees grows to the huge size a distance to 8 to 10m

Before planting it is necessary that planting place is marked 1msize pits are made well in mid summer kept in shady place.

In each pits 3 to 4 baskets of Farm Yard Manure are mixed withdug soil and filled .


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Beginning of winter the plants are planted in the centre of these

pits and staked properly.

IRRIGATION:- Taxus tree is hardy in nature and stand very well against

drought.

The crop shall be benefited by giving 2-3 irrigation at the time of

full bloom

S NO PEST AND DISEASE HAS BEEN REPORTED

TILL NOW


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HARVESTING & STORAGE:-

Taxus leaf contain higher taxol content than

bark.

Growing season is from April to October and

cut off isbanned.

Rest of the years leaves are collected.

Snowfall during winter hamper extracion.


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MARKETING AND TRADE:-

The cost of 1.7g Taxol is about 3 lakes in the

international market.


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ECONOMICS& CULTIVATION:-

The economics worked out are subject to fluctuation

depending on time and place.

On the basis of prevailing market condition the net profit

of about 4000/ha rupees in a five year old crop can beobtained.


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RECENT DEVELOPMENT INCULTIVATION OF

TAXUSBACCATA:-

PLANTCELLCULTURE:- Callus culture were started from needles and young

stems.

Wash with Tween 80 solution,plant samples weresterilized by immersion in a 70% ethanol and 2% sodiumhypochlorite solution.

Rinse with sterile water,aseptically cut into explants(partfrom plant) of about 5mm in length.

Then place in agar medium at 300k in the dark. The calli initiated on the solid medium were suspended

in a 100ml of the liquid medium and reciprocally shakenat 100strokes/min at 300k in the dark.


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To examine whether cells excrete paclitaxel into a culture

medium,suspensions of initiated callus were created in 30 ml of

liquid medium by inoculating 0.5g and 0.3g of callus tissue. Paclitaxel was detected in the medium after 2days,and its

concentration increased over time.

Paclitaxel concentration in the cells was about 10times

higher(10-15mg/kg of the wet cells) than that in the medium,thevolume fraction of cells in the culture was only 1%.

Thus 70%-80% of paclitaxel produced was released into the

medium, indicating that paclitaxel is excreted by living cells.

Paclitaxel concentration was measured in the medium

containing 2ppm naphthalene acetic acid.

Confirmation of paclitaxel was done by LC-mass spectrometry

and by immunological assay using an antibody to paclitaxel.


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PERFUSIONCULTURE:-In a 300ml Erlenmeyer flask with a mesh net cell separator shaken at 100strokes /min at 300k in the dark.

Fresh medium was continuosly fed into the flask using a peristaltic pump.

The medium was pumped out of the flask through a cell separation unit withnylon mesh net with 0.15mm openings, maintaining a constant culturevolume of 100ml.

About 1.5g of wet cells were inoculated into liquid medium after subculturing

for at least 5min on solid medium.The higher dilution rates inhibited cell growth, presumably because someconditioning factors that stimulate growth were released into the medium

The concentration of the factors were decreased by high dilution rates.

Increased dilution decreased cell growth after 19days of perfusion culture.

Inoculum size was 1.5g of wet cells in a 100ml medium.Zero value indicatesthat batch process was used.

Paclitaxel production was enhanced by higher dilution rates in perfusionculture compared with batch cultivation.

The total amount of paclitaxel secreted into the medium during 11days ofcultivation reached up to 0.38mg,


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And the fresh cell weight in the flask at day 11 of cultivation was 1.8g at a

flow rate of 90ml/day.

Considering the dry cell to wet cell weight ratio, the specific productivity of

paclitaxel released into medium was 3.2mg/g of dry cells by the 11daycultivation.

It has been reported that paclitaxel content in the cultured cells of taxus

species are 0.2mg/g of dry cells than that content in intact natural tissues is

0.5mg/g of dry cells.

Results suggest productivity was enhanced by continuous removal of the

secreted product through continuous cultivation.


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LONG TERMPRODUCTION:-Continuous paclitaxel production that used immobilized cells.

The production was conducted under same conditions used forperfusion suspension culture.

Cells were trapped in calcium alginate gel particles by drippinga concentrated cell suspension mixed with sodium alginate intoa CaCl2 solution.

The immobilized cells had a diameter of 5mm.The time course of paclitaxel production for immobilized andfree cells was compared.

The changes in paclitaxel concentrations in flow out mediumswere similar for both immobilized and free cells.

Paclitaxel concentration in the flow out medium was 0.2mg/l formore than 40 days using immobilised cells.

If the average residance time for one culture at day 16, but nochange in production profile.


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Paclitaxel production by cultured yew cells was controlled by its

concentration in the surrounding medium.

The results suggest that for better productivity ,a high dilution

rate of the medium is important.

For economical production at high dilution rates, the medium

must be reused after removing taxanes.

The system constructed composed of 2 sub systems ;the

production system, using yew cell immobilized particles,the

seperation system,for taxane removal using extraction or

adsorption.

The medium is recycled between the two subsystems.

Repeated use of the culture medium for more than 2 week in a

small scale test.

This process is now being scaled up for the mass production of

paclitaxel in the near future.


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MPH TAXUS_BACCATA

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