GRYF XBase bioSTAR 1

All the elements used for the production were selected with great care and effort to ensure maximum reliability, long service life and ease of use. All the constituents used, their functions, incl. the function of the complete equipment is under strict control.

Overview

Engaging the BioSTAR Magic Fermenter is simple and clear. All passes to the fermenter are in the hood, the connecting cables are connected to the fermenter panel the shortest way and do not prevent the user from operating. Whenever a fermentr vessel can be removed without having to dismantle accessories. Dosing pumps are part of control panels (without external connecting cables). The connection of the pumps with the fermenter vessel and the reagent containers is made by silicone sterilizable and uncoated tubing. Connecting a computer with a fermenter panel is done with one signal cable (RS485 + USB).

Universal use

Like most GRYF products, the fermenter XBase bioSTAR is a modular system.

Is possible:

• arbitrarily change the number of combined passes to another incl. changes in diameters
• add additional dosing pumps, combine their performance (volume / time, dose size)
• measure other electrochemical or physical quantities, incl. composition of the output gases
• to change aeration elements and homogenisation (stirring)
• change from rotary homogenization system to linear with adjustable stroke, change of mixers behind actuator (economical homogenization)
• measurement of the fermenter level by a capacitive sensor positioned outside the vessel

The fermenter can be used for batch and continuous cultivation.

GRYF's experience with the development of special single-purpose devices opens up new possibilities for increasing the utility value of a fermenter and a high technical level. 

Operator comfort

All processes can be controlled directly from the PC either in automatic or manual mode.

The fermentation process can be monitored outside the workplace within the local computer network or the Internet. Based on the requirements, the existing system can be supplemented by additional hardware and software upgrades. Some functions, such as venting and pumping liquid into the dosing pump, can be done manually in a pre-set mode.

GRYF bioSTAR fermenter description

The fermenter consists of several basic parts:

• Sterile fermenter vessel incl. accessories
• Heating and cooling system
• Stirrer with drive unit
• Two control panels with pumps and other accessories
• Compressor
• PC inc. control software
• Measuring system

All the controls are located on the front sides of the control panels.

On the back of the control panels are the measuring and automation elements, accessible under the entrance door (only for service). All inlets and cable connections are connected from the back side.

Fermenter vessel - the vessel is made of the borosilicate glass, cylindrical, straight, rounded edges. The straight bottom provides a sufficient heat transition for heating and cooling the fermenter. The lid is made of stainless steel, the seal between the vessel and the stainless steel lid is silicone rubber O-ring. Hermetic closure is provided with an original stainless steel buckle supplied by the vessel manufacturer.

The vessel size (the volume of the fermenter vessel) is determined by its base - 112 mm diameter and 280 mm maximum container height. The basic version uses a container with a usable volume of 500-1000 ml (net volume 1260 ml). The vessel height is 175 mm. The container has a volume mark of 250ml.

The fermenter lid is equipped with stainless steel or PVDF passages with a silicone rubber sealing element and a pull-out tightening nut. In the event that the passages are fitted with accessories, they are sealed with seals. The tightness of the passages is ensured with a large margin of pressure up to 8 (stainless) / 4 (PVDF) bars. All elements of the passages can be permanently loaded up to 150 °C.

The fermenter lid has a total of 6 passages.

3 separate passages "A1, A2, A3" for fitting and fixing accessories with a diameter of 12 mm.

• electrode for pH measurement
• electrode for pO₂ measurement
• probe for measuring the turbidity

After inserting the probe and placing it at the right level, its body is locked by a mask that presses the silicone sealing element to the probe and closes the hermetic opening.

• drainage of condensate return gases

1 combined passage "C1" 6x 3.5 mm (5 stainless cannulae for liquids, 1x aeration)

• feeding
• vaccination
• collection, sampling
• dosing antifoam
• aeration

1 combined passage "C2" 2x 5(4) mm

• temperature sensor
• 1x reserve

All inserted accessories are secured in the passage by tightening the swivel nut.

The magnetic coupling divider element that ensures the hermetic drive of the stirrer is in the middle of the lid.

Complete tank assembly incl. passages and the separating element of the magnetic coupling is autoclavable.

Cannulae

They are made of stainless steel thin-walled tubes of the diameter 3-3.5 mm. At the end they are finished with an olive with a plug-in nut for connecting the hosepipe. Material stainless steel/plastic resistant to temperature min. 135 °C. Upon user request, the dimensions of the cannula can be adapted to its needs.

The cannulae are autoclavable together with the fermenter vessel.

K1 - longest cannula ending at the bottom - sampling (harvesting)

K2 - cannula terminating at surface level - vaccination

K3 - cannula terminating above the surface level - defoam

K4 - cannula end at the bottom - reagent dosing for pH adjustment

K5 - optional - fattening

K6 - optional - air supply

Tempering

Tempering is ensured by a powerful Peltier's segment. Possible temperature range is 0-70 °C. Operating temperature range 10-60 °C, setting at 0.1 °C. The temperature difference in the cooling range compared to the ambient temperature at full power is Δt max. 19 °C. In the cooling process, heat insulating material (foam ring) can be used on the fermenter vessel to accelerate tempering to temperature stability. The excess heat produced by the Peltier's cell is drained by the active venting of the back of the fermenter base. Temperature sensor for temperature control is located along with other sensors inside the fermenter (passage C2). Stainless steel design. Autoclavable.

Both the set and actual temperature is displayed on the computer monitor.

Mixing - homogenization of fermenter content

The mixing device is a mechanical - stainless steel shaft on which the turbine can be placed with radial and axial pressure on the liquid. Turbines work in the laminar flow area. Turbines and locations depend on the user's decision. It is possible to place up to three turbines on one shaft with full volume (1000ml). Engine power of the agitator increases provided to the turbine depending on the diameter and speed.

The empirical formula for indicative calculation of the input power for laminar mixing of the liquid:

                                        N = A . d3 . n2 .  η

N = engine power of the agitator,  A = agitator type coefficient, d = agitator diameter,  n = rotation number,      

 η = liquid viscosity

Maximum diameter with one turbine (standard geometric shape) is 70 mm at 200 rpm. The agitator can be complemented by new user-designed geometric shapes of the turbines (complex shapes).

All mixing parts placed inside the fermenter are autoclavable.

The agitator drive unit is a DC motor with planetary gearing. This motor is controlled by PWM (pulse width modulation) in the range of 0-240 rpm. Connecting the drive unit to the agitator is via a magnetic coupling. The torque is 1 Nm at 200 rpm. With lower speeds due to the control electronics torque increases. The drive unit is equipped with an encoder to accurately monitor the set speeds.

The customer may choose a different range of stirrer speed after consulting the supplier.

The agitator speed is displayed on the computer monitor.

The agitator drive unit is located on the main shaft and can be ejected at any time and disconnected from the magnetic coupling. The connection of the magnetic coupling with the shaft of the drive unit is a mechanical quick-coupling.

Aeration – The source of air is a small, unleaded, quiet laboratory compressor (noise below 50 dB). A bacterial filter for solid particle filtration and possibly a small amount of suspended liquid in the gas is located at the outlet. Design AA: Efficiency at filtration level 0.01m: 99.9999 +%                                    

The air dosing volume is controlled by a proportional valve with a mass flow meter. A flowmeter with a control valve is part of each fermenter. User-adjustable scope range from 0 -5 l/min. is controlled in the range 0-100%. The dose is continuously controlled according to the measured and required partial pressure value pO₂ or manually. The flow is recorded by the measuring system and is displayed on the monitor.

The aeration element is of a standard design - a circle or nozzle membrane located at the bottom of the vessel. Material stainless, autoclavable.

The aeration system can be supplemented with new user-designed aeration elements.

The connection of the clean air source with the fermenter vessel is with a silicone hose. For preventive reasons (to prevent possible contamination), a disposable antibacterial filter should be added to the fermenter's inlet.

Dosage - Peristaltic dosing pumps

WELCO - Japanese manufacturer. Precision pumps with support track, with two or four rollers, with a sufficient flow rate controlled by a stepper motor. The stepper motor allows you to dispense even very small doses. Dosing pumps are built directly into the fermenter panel. There is a large selection of hoses of different diameters and chemical resistance (see Table). The pumps can be set to two basic modes in automatic mode:

1. dosing ranging from 0-100% in a preset range from the max. range of the PC controlled pump
2. "dose" mode - set repeatable dose (eg, dispenser dosing, etc.) controlled by PC

Dosing pumps are proportionally controlled from the PC based on the evaluation of the control system. One turn = 12,800 steps.

Dosing pumps can also be operated manually (button) with the function of venting and sucking liquid. With the mechanical switch on the panel, 10 power levels (rpm) can be set for manual operation. Running manual operation takes precedence over automatic control from the PC and is independent of the state of automatic fermenter control.

Each pump can be assigned an "Integrator" feature (a new XBase XBC software feature). This function will ensure that reagent consumption is monitored over a given time period.

By monitoring the consumption of pH-adjusting reagents, it is also possible to monitor the growth of cultivated micro-organisms indirectly according to the culture and replace the turbidity measurement.

The drive tubes of the metering pumps are autoclavable.

Dosing pump functions

DP1 feed

20 rpm, 12800 steps/rev

Tube type: PHARMED BPT inner diameter 3,2 mm

Max. dose/min: 10 ml/min

Dose/rev: 0,5 ml

Preset: 10 ml/min

DP2  pH adjustment

20 rpm

Tube type: according to specification

Max. dose/min

The dose per step

Advantage: according to user input

DP3  dose antifoam

20 rpm, 12800 kroků/ot.

Tube type: according to specification

Max. dose/min

The dose per step

Preset: Tube selection and settings will be tailored to user requirements

DP4  reserve

DP5  collection, sampling

0-150 rpm, 1600 steps/rev

Tube type: according to specification

Max. dose/min

The dose per step

Preset: Tube selection and settings will be tailored to user requirements

Vacuum pump

Starting is done manually, the suction control is controlled by a control valve. When extracting liquid from the fermenter into a collecting container, the level must be monitored to prevent liquid from entering the vacuum pump.

Level measurement

Level monitoring is performed by stainless steel electrically conductive cannula located in the reactor. The lowest cannula (K1) is one contact of "conductive electrode". The second contact forms a cannula (K2) located at the height level. The end of the third cannula (K3) is located above the surface, at an height where the foam is assumed to increase. The used cannulas are connected by a common cable to the reactor panel. The level signal is passed to the PC for evaluation.

Feeding, automatic maintenance levels

These functions are directly related. Unless otherwise stated, the level of the fermenter is maintained replenishing the nutrient solution. Based on the conductivity probe information, the dosing pump is controlled and the level is added to the given value. This function works in ON/OFF mode.

Automatic antifoam

Similarly to maintaining the level, a third conductivity probe is used to measure foam formation. Upon reaching the foam level to the conductivity electrode, a signal is evaluated and a dosing pump is automatically switched on, which delivers a pre-defined volume of the antifoam to the reactor. If a suitable selective defoamer is used, the foam drops and the signal is inactive. Upon further raising of the foam, the process is repeated. Antifoaming can also be triggered manually by holding down the button on the appropriate dosing pump. This action does not affect on the process of the automatic dosing.

Harvesting, sampling

The fermenter's equipment allows several ways of harvesting cultivated media.

The sampling is performed from the lowest cannula.

1. using a peristaltic pump
2. using the vacuum source (vacuum pump) that is part of the fermenter equipment
   • collection into an auxiliary container with subsequent transfer to the target container
   • collection directly into the sampling target container
3. Manual collection  into an auxiliary container using a syringe

Inoculation

Inoculation can be performed through the cannula 2, which end is located on the surface.  The cannula is fitted with a silicone seal to prevent contamination. This bulkhead could be overcome by the injection needle. Once the vaccination has finished, the cannula should be closed again.

Gas outflow from the reactor

Gas outflow is provided by a separate grommet in the fermenter lid (B1). A small glass cooler is provided at the outlet to ensure condensation of vapors and return of condensate to the reactor volume. It is possible to connect a cold water source to the cooler. The surface of the cooler walls and the ambient temperature may be sufficient for the normal operation of the fermenter. The outlet of the cooler must be fitted with an antibacterial filter.

The output can be used to measure the output gas composition (O₂, CO, CO₂, CH₂, H₂S, etc.). Gas composition measurement is not part of the basic equipment (extra equipment).

Measuring system

The hardware and software of the XBase XBC measuring system are used for the measurement of electrochemical and physical quantities, including regulating actuators.

pH measurement and reagent dose control for pH adjustment

The pH electrode is placed in the reactor and fixed in a stainless steel lid. The electrode connector is connected to the fermenter panel. The electrode signal is processed by the XBase XBC measurement module (measuring heads) located in the fermenter panel.  The measured pH information is passed to the PC where the information is evaluated and the action is performed by the pH adjustment pump DP2 according to the set criteria.

The type of pH electrode used: Mettler Toledo InPro3253/120/Pt1000 -  the electrode is sterilizable and autoclavable.

Measurement of partial pressure pO₂ and aeration control

The electrode for measuring pO₂ is placed in the fermenter in the same way as the pH electrode.

The signal of the electrode processes the XBase XBC measurement module (measuring head) located in the fermenter panel. Information about the measured concentration of pO2 is transmitted to the PC, where the action is evaluated and according to the set criteria.Airflow control is via a proportional valve with a mass flow meter.

The type of the electrode used: Mettler Toledo InPro6800/12/120 - the electrode is sterilizable and autoclavable.

Measurement of turbidity

The Mettler Toledo stationary instrument is used for turbidity measurement.

Transmitter typ TRB 8300

Probe InPro8200/S/Kalrez-FDA/120 3.1

The cloud measuring probe has a diameter of 12 mm and can be connected to the fermenter via a universal access passage (eg A3). If the probe is not placed on the fermenter from the beginning of the process, it can be inserted into the fermenter using the above-mentioned passage. Even though the electrode is sterilizable, there is a risk of contamination of the fermenter content when opening the passage. The second option is to take a sample in which the measurements are made. With a permanent turbidity probe connection, the probe can be connected to the XBase XBC measurement system as other probes and electrodes, using the full comfort of this measuring system, with the ability to control the density of the cultured biomass.

Software

The XBase XBC measuring system with superstructures (Client - Remote Surveillance and Graphs - Working with Charts) is used to control the fermenter in real time. Installation CD or USB disk with the appropriate software is supplied to each fermenter.

Software list:

• XBase XBC version 0.93 and higher 1 license
• XBase XBC client version 1.0 and higher multi-license
• XBase XBC graphs version 1.4 and higher multi-license
• Text files user manual for the software
• Text files operating and maintenance instructions, service instructions

Although the existing XBase XBC software seamlessly manages the fermenter's control within a given range, it will be complemented free of charge by the "Integrator" and "Batch" functions (a function for improved control and control of dosing pumps).

Since the basic software is still under development, a higher version may be provided.

The installation software for PC is also included.

A list of displayed monitor quantities

On the taskbar conditionally with settings unit

Fermenter temperature measurement Set temperature °C

pH measurement Set pH pH

pO2 measurement Set pO2 mg/l

Air flow - aeration ml/min            

Turbidity FTU

Agitator speed n/min

Computer assembly

Renowned computer manufacturer AT Computers has been chosen as a supplier. This company supplies computers for its subsidiary Autocont and many others throughout the Czech Republic. Thanks to this vendor, we can guarantee high device reliability and fast response to hardware and software issues. 

Execution

InWin EA-009 midi case black / silver (dimensions: 190px x 445h x 415v). motherboard with P43 chipset.

Keyboard USB

Optical mouse USB

RS485 card for communication and control of the fermenters

The following software is included in the delivery of the PC:

Original Microsoft Windows 7 Professional CZ

Office Home and Business 2010 DOEM CZ

Monitor

22" LCD Acer E220HQb -50 000:1,5ms,Full HD, black

Barva case: black

Autoclaving

Autoclaving is done by standard procedures. It is necessary to ensure that all unattached passes through the plugs, electrodes and sensors are provided with the protective cover of the connectors, to dismantle the components that are to be autoclaved.

The manufacturer reserves small dimensional and layout changes to the instrument.