he strain used in this invention has complex nutritional needs.

Therefore, the growth medium must

contain between 12 and 13 % of glucose, 0.25% of ammonia and phosphate, and B complex
Vitamins. In order to satisfy these needs, after the dilution of the molasses, a solution of
(NH4)2HPO4 has to be added, to suppress the growth medium lack of these two components.

The growth medium contains a wide variety of remaining cells and spores from its constituting
elements. These must be eliminated prior to inoculation, in order to maximize the fermentation yield.

The sterilization occurs continuously and takes place in heat exchangers, with the shape of concentric
tubes, where steam or overheated water at 140 °C passes trough the external tube and the growth
medium passes trough the internal tube. The hold-up is between 30 and 120 seconds depending on
work rate. The medium must then pass trough a cooler, in order to reduce its temperature to match
the fermentation temperature.

To reduce energy costs with heating and cooling the medium, un-sterilized medium (cold) is crossed
with sterilized medium so that the last one can be cooled to the fermentation temperature.

This type of sterilization has the advantage of reducing the sterilization cycle time, easy control and
productivity increase, without the destruction of the existing nutrients in the solution. During the
sterilization process it is common to observe the formation of foam. Adding a tensioactive compound
such as Tween 80 in a concentration of 0.001 % can minimize this effect.

The sterilization of the reactor and of the filtration systems is made by direct steam injection; this is
possible because the filtration membranes are thermoresistant.

In order to increase even further the process productivity, the fermentation takes place in a
continuously agitated reactor, like a chimostat, with a high cell concentration. This system avoids,
simultaneously, substrate inhibition - since the substrate is continuously supplied into the fermenter -
and product inhibition - since the lactic acid is continuously removed from the fermenter.

The high cell concentration is obtained trough cell immobilization. The immobilization is achieved by
restraining the cells to a confined space using a physical barrier. The physical barrier used in this
invention is cell recycling trough ultrafiltration membranes. This method has several advantages,
mainly in product productivity increase, and in high fermentation yield. It also presents low energy
costs and advantages in operation safety and stability of the production system.

Furthermore, this system prevents many of the mass transfer problems, caused by diffusional
problems, from occurring and also prevents contamination of the fermentation broth, where the final
product can be found, as well as contamination of the fermenter, where the fermentation is occurring.

At the same time, this system allows operating at high dilution rates, without system collapsing,
phenomenon known has wash-out. The high dilution rates at which the system operates is another
very important factor that contributes to the productivity of the system. All of these factors combined
make this a viable and profitable system.

For the fermentation process to be initiated, a pre-inocule must be prepared, with 10 % of the
reactor's work volume. This can be prepared in smaller reactors. Moreover, the fermentation process
must begin with a discontinuous phase of about 8 hours in order to obtain biomass.

After this discontinuous period, the bioreactor is ready for full functioning. A general diagram of the
fermentation and purification process can be observed in the process description diagram. Probes
control the fermentation conditions continuously. The information is analysed so that the conditions
are stable, especially regarding temperature, pH and cell concentration. The pH is controlled by
addition of concentrated NaOH (recovered from following process steps).

The purification of the Lactic acid, that exits the fermenter in the form of Sodium Lactate, due to the
added NaOH, is initiated in the ultrafiltration membrane system used to maintain the high cell
concentration. These ceramic membranes retain bacterial cells and other macromolecules, which are
then reintroduced in the reactor. The fermentation broth containing the lactic acid is conducted to
further purification.

The next purification step takes place in a nanofiltration system. This system contains nanofiltration
membranes with an exclusion limit between 0.5 and 5 nanometers. This way all of the contaminants
that are bigger than the lactic acid are eliminated.

The separation of the sodium lactates in lactic acid and NaOH, and the consequent concentration of
the lactic acid is achieved through bipolar electrodialysis. The resulting NaOH is reused for pH
control.

Electrodialysis is an electrochemical process that allows the separation, purification and concentration
of ionic substances, by applying an electrical potential difference. \Ve is the use of the bipolar
membrane that allows the purification and concentration to occur simultaneously. By using bipolar
electrodialysis the entire separation and purification process is simplified, eliminating by-products and
avoiding the use of organic solvents.

From the electrodialysis process result two fractions, one containing lactic acid and residual sodium
lactate, and the other fraction, containing NaOH. The residual sodium lactate fraction can be reduced
by increasing the number of bipolar membranes used.

The lactic acid purified in this process in intended for the production of polylactic acid (PLA). PLA is
obtained by polymerisation (ring opening), of a cyclic dimmer of lactic acid (intermediate) without the
use of organic solvents.

The lactic acid, produced, purified and concentrated in the previous steps is transported to a CSTR
(continuously stirred tank reactor), where it will be dehydrated and pre-condensed. The reactor is
equipped with a stirrer, a condenser and temperature control devices, important to assure that the
reaction occurs for 2 hours at temperatures between 160 and 200 °C, at atmospheric pressure. The
effluent water contains only residual lactic acid, meaning that it is not considered as a dangerous
effluent and therefore does not require any treatment.

From the first polymerisation process lactic acid dimmers and oligomers are obtained, which are then
separated, purified and concentrated by distillation in a distillation column. The effluent resulting from
this process is recirculated and reintroduced in the first polymerisation reactor (CSTR 1).

The concentrated dimmers and oligomers are then transported to a second reactor (CSTR 2) where
the polycondensation reactions will occur. The reaction time for these reactions is 20 hours at
temperatures between 160 and 200 °C, and at reduced pressure around 0,2 - 20 mmHg, in the
presence of a catalyst, iron lactate, used in a concentration of 0.005-0.5% (p/p), regarding the initial
lactic acid. The polycondensation reactions should take place, maintaining the initial conditions, until
the desired molecular weight is achieved, around 25000 - 30000 kDa. The condensation step can be
performed in several steps, lowering the pressure gradually.

The yield of the polycondensation reaction is around 80% (p/p), and the yield in dimmers and
oligomers around 20%. The polylactic acid is collected and the resulting dimmers and oligomers are
transported to a fractional distillation column, where these dimmers and oligomers are purified at
temperatures between 144-170 °C and a pressure between 0.5-0.8 mmHg. From this fractional
distillation, purified dimmers are obtained which can be recycled and reintroduced in the
polycondensation reactor (CSTR 2), and pure lactic acid residues and water, which are recycled and
reintroduced in the first polymerisation reactor (CSTR 1). Since the non- condensate fractions are
recycled, the overall yield is close to 100 %.

COMMERCIAL APPLICATIONS AND FUTURE PERSPECTIVES

The production of organic acids and aminoacids with low production costs is a long pursued objective
in several industrial applications: food industry, animal feed and biodegradable plastics, in the case of
lactates.
This polymer has several advantages comparing to common petroleum derived plastics:

• Biodegradable, which means that microorganisms are able to alter the plastic properties and modify
its chemical structure.

• Biocompatible, which means that these plastics are, in normal conditions, not rejected by biological
cells or tissues? • Produced from a renewable resource (an industrial residue), and not from
petroleum- derived products.

• 100% recyclable: Trough hydrolysis the lactic acid can be obtained and reused for a different
application or it can be merged to produce another object

• No organic or toxic solvents are used in the production of these biopolymers.

• They can be incinerated emitting only CO2 (previously removed from the atmosphere) and H O.

PLA can be produced with a wide variety of properties and characteristics, because the lactic acid
molecule is a chiral molecule with two asymmetry centers and therefore three possible enantiomers
derived from this dimmer. This way controlling the stechiometry of the lactic acid dimmer formation, it
is possible to obtain different mixtures of lactic acid containing the L and D or meso-lactate forms.

These different mixtures result in different properties such as strength, stiffness and others. PLA is
often compared to PET (Polyethylene thereftalate), due to the enormous similarity between the
properties of the two compounds.

These polymers are resistant to fats and are an effective barrier against odours and flavours. They
have a better resistance to heat variations then the common polyolefins.

PLA can be used for the same purposes as normal polyesters. The main applications are:

• Product wrapping, for instance candy or caramels

• Natural textile fibber

• Wide array of medical applications, such has bone implants, biodegradable stitches or
biodegradable capsules or vectors for pharmaceutical and chemical compounds.

• Besides the applications referred above PLA has many other different applications in the form of
fibber, resin or thermoplastic. Future perspectives are very positive for PLA. The decrease in the price
of production is increasing the range of applications and the level of substitution of petroleum-based
plastics for biodegradable plastics such as PLA will increase significantly in the forecoming years.

The technological developments in recent years can drop the production price of PLA production to
levels close to the price of common plastics, making PLA more and more competitive. Moreover,
Governmental pressure, mainly in Countries in the European Union and Japan, is forcing package
producers to find solutions for the environmental problem caused by the enormous accumulation of
urban solid residues. All of these arguments support the use of PLA as a viable alternative to common
plastics.

DISCLOSURE OF THE DIAGRAM OF THE INVENTIVE PROCESS

The diagram represents the continuous process for the production of polylactic acid from sugar
molasses.
The entrance flow (1) (constituted by fresh broth previously prepared), passes through a sterilizer and
then through a cooler (2) (where it is sterilized and cooled to the fermentation temperature). It then
enters in the fermentation tank (3) in witch sugars are fermented to lactic acid.

The temperature inside the tank is held constant with circulating water. Into the fermentator enters a
sodium hydroxide flow (4) from the electrodialysis system (5).

The exit flow (6) feeds an ultrafiltration system (7) in witch cells are separated from the fermentation
broth. Cells can or not (in case of purge), re-enter the fermentator through flow (8). The fermentation
broth, containing the sodium lactate flows (9) to a nanofiltration system to separate macromolecules.
The permeate flow (11), containing the sodium lactate and other solutes smaller than the cut-off of the
filtering membranes, enters the electrodialysis system (5) to concentration and purification of the lactic
acid; The held can or not, be recirculated to dilute the entrance flow (1), through flow (10).

From the electrodialysis system (5) result three different flows: one containing NaOH (4), another the
impurities (12) and the last one, lactic acid (13).

Flows (13) and (14) enter the CSTR (16) - flow 14 comes from the fractionated distillation (15). From
reactor (16) exits, by evaporation, a flow (17) containing water and traces of lactic acid, and another
one (18) containing the dimmer and other oligomers that are directed to a distiller (19).

From this distiller come out two flows (20 and 21) that mix with flow (14) and the polycondensation
reactor (22), respectively. From this reactor (22) come out a flow (23) containing the final product
(polylactic acid), and another one (24) containing dimmers and all lactic acid oligomers that can be
separated by fractionated distillation (15).

Dimmers can be directly polymerised in the casting box (25) by adding a catalytic octanoate (between
180 and 195°C) and impurities are recirculated to flow (14) to a new polycondensation

US20050119448A1