WELCOME

BIOSOLUBILIZATION IN FLUIDIZED BED BIOREACTOR

PRESENTED BY, GUIDED BY,HARITHA K M LEKSHMI MISSROLL NO 7111 ASST PROFESSORBT AND BCE BT AND BCE

SREE BUDDHA COLLEGE OF ENGINEERING,PATTOOR

BIOSOLUBILIZATION OF MINERAL

INSOLUBLE PHOSPHATES BY

IMMOBILIZED MICROORGANISMS

IN FLUIDIZED BED BIOREACTOR

CONTENTS I n t r o d u c t i o n Ro l e o f P h o s p h o r o u s I n P l a n t s P h o s p h o r o u s C y c l e P h o s p h a t e S o l u b i l i s i n g M i c r o o r g a n i s m s Ty p e s o f P h o s p h a t e S o l u b i l i s i n g B a c t e r i a P h o s p h a t e S o l u b i l i s i n g Fu n g i S o i l M o b i l i z a t i o n A n d I m m o b i l i z a t i o n B y B a c t e r i a M e c h a n i s m s o f P h o s p h a t e S o l u b i l i s a t i o n S e l e c t i o n o f P h o s p h a t e S o l u b i l i s i n g M i c r o o r g a n i s m s P h o s p h a t e S o l u b i l i s e I s o l a t e d Fu n g a l S t r a i n Fo r m i n g C l e a r Z o n e I m m o b i l i z a t i o n Fu n g a l B i o m a s s I m m o b i l i z a t i o n Re p e a t e d B a t c h O p e r a t i o n U s i n g Fr e e a n d I m m o b i l i s e d Fu n g a l B i o m a s s r e s u l t s M o d e o f A p p l i c a t i o n O f P h o s p h a t e S o l u b i l i z i n g M i c r o o r g a n i s m s A d v a n t a g e s D i s a d v a n t a g e s C o n c l u s i o n

INTRODUCTION The world population increasing day by day.

Hence there is need for plenty of food crops to meet the requirement. Crops need several nutrients to reach their maximum potential yield.

INTRODUCTION(CONT…)

Phosphorus is an essential macronutrient for plants.

Phosphate fertilizers is easily precipitated into insoluble forms.

Solubilisation of inorganic phosphates by microorganisms.

These microorganisms was entrapped in alginate and polyacrylamide gels. Used for solubilizing mineral insoluble phosphates in fluidised bed bioreactor. ADVANTAGES Reusability of the same biocatalystControl of reactions No contamination of products

INTRODUCTION(CONT…)

ROLE OF PHOSPHOROUS IN PLANTS

It plays an important role in metabolism of crop plants. About 10 to 25% of fertilizer phosphorous is acquired by plants. It is the constituent of various cellular functions or activities such as cell division ,development ,photosynthesis , breakdown of sugars etc. Plants deficient in P, shows retarded growth.

PHOSPHOROS CYCLE Phosphorous is taken up by the plants from soils and it is consumed by animals. Return to soil as organic residues. Conversion of organic phosphate to inorganic phosphate is known as Mineralization

PHOSPHATE SOLUBILIZING MICROORGANISMS

In soil phosphate solubilizing bacteria constitute 1 to 50%,while phosphate solubilizing fungi are 0.1 to 0.5%. Phosphate solubilizing activity is greater for fungi than bacteria.1 Repeated sub culturing2 Produce more acids 3 Traverse more distances

TYPES OF PHOSPHATE SOLUBILIZING BACTERIA

BACILLUS

PSEUDOMONOS

PHOSPHATE SOLUBILIZING FUNGI

Aspergillus awamori Aspergillus conidia Aspergillus colony

PHOSPHATE SOLUBILIZING FUNGI

Aspergillus fruiting body Penicillium Trichoderma viridi

SOIL PHOSPHOROUS MOBILIZATION AND IMMOBILIZATION BY

BACTERIA

MECHANISMS OF PHOSPHATE SOLUBILIZATION

Phosphate solubilizing microorganisms convert insoluble phosphates into soluble forms . Through the processes of acidification, by the production of organic acids, production of acid and alkaline phosphatases. These organic acids can either dissolve phosphates as a result of anion exchange or can chelate Ca,Fe or Al ions associated with phosphates.

SELECTION OF PHOSPHATE SOLUBILIZING

MICROORGANISMS Grow bacteria/fungi in specified media containing insoluble phosphate. Incubate and observe the clearing zone around colonies. Quantify the phosphorous dissolved by the specified organism. Select efficient culture based on above criteria.

PHOSPHATE SOLUBILIZER ISOLATED FUNGAL STRAIN

FORMING CLEAR ZONE

IMMOBILIZATION Conversion of inorganic compounds to organic compounds by microorganisms. It is prevented from being accessible to plants. Opposite to mineralization

DIFFERENT METHODS OF IMMOBILISATION

FUNGAL BIOMASS IMMOBILIZATION

ENTRAPMENT IN CALCIUM GEL: 100ml sterile sodium alginate + 2g fungal biomass Mixture was extruded into 150 mM Calcium Chloride, forming beads of 5mm diameter

ENTRAPEMENT IN POLYACRYLAMIDE GEL: 2g fungal biomass + 78ml Tris-HCL buffer+20 ml bisacrylamide solution+1ml ammonium persulfate Polymerisation is initiated adding 100μl of N,N,N’,N’- tetramethyl-ethylenediamine

FUNGAL BIOMASS IMMOBILIZATION (CONT...)

FLUIDISED BEDBIOREACTOR

Small particles are suspended Upward motion of reacting fluid Used when intimate content between the solid particles and fluid is required. Solid particles could be flocculated cells Fluid could be fermentation broth.

FLUIDIZED BED BIOREACTOR USING FREE AND IMMOBILIZED

FUNGAL BIOMASS Fluidized bed bioreactors composed of 500 ml conical flask containing the immobilized fungal biomass Suspented in 200 ml of NBRIP liquid medium and supplemented by 5g/l of DCP Placed in a rotary shaker at 25°C Fluidisation was assured by stirring at a rate of 120rpm Phosphate solubilisation rate was followed according to time in bioreactor. Supernatants was analysed spectrophotometrically to determine amount of soluble phosphorous

REPEATED BATCH OPERATION USING FREE AND

IMMOBILISED FUNGAL BIOMASS

Longevity of solubilisation activity of the immobilised fungal biomass was investigated in repeated batch solubilisation tests.

RESULTS

MODE OF APPLICATION OF PHOSPHATE SOLUBILIZING

MICROORGANISMS

Soil application method

ADVANTAGES Greater population Faster action Withstand low moisture content Improve plant growth and yield Increaasing fixed phosphorous in soil

DISADVANTAGES Phosphate solubilizing bacteria not only release phosphate but also biological components such as auxin,gibberellic acid as well as vitamins The poor survival and colonization Strong competition from native microbes Availability of inadequate nutrients

CONCLUSION Bioconversion occurs at a low temperature and is more selective to phosphate extraction than conventional processes. Biophosphoros fertilization is an achievable goal that tends itself well to the sustainable agricultural production. Apart from phosphate solubilizing abilities, some of these microorganisms can benefit plant growth by several different mechanisms such as enhancing nitrogen fixation, plant hormone production .

REFERENCES Zeroual Y, Chadghan R, Hakam A, Kossir A (2012) Biosolubilization of Mineral Insoluble Phosphates by Immobilized Fungi (Aspergillus niger) in fluidised bed bioreactor. J JBiotechnol Biomaterial s6 dx:htpp//.doiorg/104172/2155-952X,s6-004 R.Jain et al./Applied Soil Ecology 46 (2010) The evaluation of free encapsuled Aspergillus awamori for phosphate solubilization fermentation and soil plant system. Applied soil ecology 46 90-94 Rashi Awasthi R Tewari and Harsh Nayyar(2011)Synergy between plants and p- solubilizing microbes in soils:Effect of growth and physiology of crops.International research journals vol.2(12)pp 484-504 Ahmad Ali Khan and Ghulam Jilani(2009)Phosphorous solubilizing bacteria: occurrence , mechanisms and their role in crop production. J.Agric.biol,.sci 1(1)(48-58) Praveen M Patil, V B Kuligod (2012)Effect of phosphate solubilizing fungi and phosphorus levels on growth, yield and nutrient content in maize.Karnataka J. Agric.sci,25(1)(58-62) Satya Sunder Bhattacharya and Soma Barman (2013)Phosphate solubilizing ability of Emericella nidulans strain isolated from vermicompost Indian journal of experimental biology.Indian Journal of Experimental Biology vol 51,October 2013,pp.840-848