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Since the purchase of the device, it has been idle for many years and passed the equipment warranty period. Due to the needs of the work, the NBS40L fermenter is required, and the NBS40L tank is a high-tech automatic fermentation equipment. So we first translated and familiarized with the operation manual, and then found the pipeline. On this basis, the equipment was successfully activated. . In the half year of using the NBS40I tank, the initial failures occurred, and new problems appeared from time to time, and we all solved them one by one. Now we think the BS40L fermenter is a very good device. It is the process of digestion and absorption of this imported equipment, and the process of discovering and solving problems is very meaningful. To this end, the experience of using NBS40L fermenter is introduced to everyone:
First, the control board failure solution <br> In the first test machine, there is a Ding temperature control failure, the temperature display does not match the actual temperature, more serious is no way to sterilize. After consulting the electrical engineer, the control board integration block was broken. After repairing, the fork is not used for a long time, and the temperature control is not allowed. The speed cannot be controlled. After the inspection, the control board integration block is broken. We found that the time of failure occurred mostly during night cleaning and disinfection, and during autoclaving operation. After analyzing several faults, we believe that the equipment has strict requirements on voltage range and fluctuation (Table 1). At present, the voltage in the plant varies greatly (Table 2), especially after the night power unit is reduced, the voltage is increased or not. Stable and easy to break through into blocks, causing malfunctions. For the voltage problem, we add a tube regulator to the fermenter. Model SH1742 (5Kv) electronic AC voltage regulator. The same problem did not occur in the next twenty batches of fermenters. If the imported equipment fails due to voltage problems, it is common that the 110V voltage is misconnected to 220v, and the fuse or other parts of the equipment are burned out. Due to the large voltage surge and breakdown of the circuit block, equipment failures rarely occur. There are no special requirements for this in the specification of the NBs40L fermenter. The same failure occurred in the original NBSsL tank. NBS40L automatic fermenter is necessary to install a voltage regulator to supply the Chinese market. To reduce the failure due to voltage changes.
Second, the fermenter enters the water, causing one of the causes of contamination
In the 27 batches of products using NBS40L cans, 6 batches of bacteria were present. After analysis, the batch number of these batches of bacteria is mainly due to the water in the fermenter during the cooling process after autoclaving, and the volume increases by about 2L. The bacteria were positive for the fermentation before inoculation. The reason for the water in the fermenter is that according to the Equipment Operation Manual, the pressure of the agitator shaft seal may be too large: the equipment requires that the shaft seal pressure be below 10 PSI, but in fact the pressure we control is 5-8 PSI, so Not the reason for the water. After repeated trials and analysis, the reason is that when the fermenter is autoclaved and sterilized, it enters a cooling state, and a negative pressure is generated due to a drop in the temperature of the tank. When the cooling temperature drops to 47 ° C, the pneumatic valve that enters the culture control state from the cooling state and the pneumatic valve of the exhaust gas are opened. Under normal conditions (can design state), the pressure inside and outside the tank will soon reach equilibrium. However, because the exhaust filter becomes wet during sterilization or clogging for a long time, the balance process is prolonged, and the condensate discharge of the exhaust filter is not tightly sealed by the pneumatic valve. The water in the condensate pipe is This continuous negative pressure suction tank increases the tank volume and contaminates the tank fluid. After finding the reason, we established a scientific operation method. On the one hand, the exhaust filter was cleaned, soaked in 75% ethanol solution, and dried before use to greatly reduce the negative pressure generation time. On the other hand, in the front of the pressure balance to the condensation water pipe (fine The arm) continuously passes through the steam, effectively preventing the condensed water from sucking up the tank; and the third periodically flushing the pneumatic crucible with distilled water to keep it clean. In the subsequent fermentations of dozens of batches, there was no problem of bacterial infection, and the results were satisfactory. It can be seen that the design of the NBS40L tank is stricter on external conditions and operational requirements.
The sterilization control of the NBS40L tank is carried out automatically. When the steam pressure is adjusted and the sterilization time and number of revolutions are set, the sterilization process (121.5 ° C) is automatically completed. But there are still two issues that need to be addressed.
(1) The heating rate is too slow or does not reach 121.5 °C. Analysis of the reasons, the steam pressure is too low to reach the requirements (less than 16 PSI). A very important reason is that the pneumatic air pressure (instrument air pressure) is not enough. The equipment requires 60 PSI, about 0.4 MPa. In fact, it can not be automatically sterilized normally at 0.4 MPa, and the required pressure range is 0.4-0.7 MPa. The lack of a pressure equalization device is a deficiency of the device.
(2) The volume of the tank liquid increases greatly. For fermenters autoclaved with steam. It is normal to increase the volume during the sterilization process, but how to reduce the increased volume is indeed an operator's experience. After many attempts, we will increase the volume after sterilization from 2 to 3L to about 1L. The reason for the increase in volume during fermentation of the fermenter is that the inoculating port, the sampling port, the inlet pipe, and the discharge port are generated during steam sterilization, and the tank temperature is generally required to reach 100 ° C. The inlet valve and the exhaust port of each nozzle can be opened for sterilization. If the temperature of the tank liquid reaches 121.5 ° C, the nozzles are opened for sterilization, and the problem is solved.
Fourth, spare parts to buy equipment <br> An important problem is to find out which are the necessary components of the equipment, which are optional spare parts (Optiona1). The price of equipment is often closely related to it. The spare parts of the NBS40L fermenter are not enough. In order to solve the problem that the acid and alkali ports are too small in the flow rate of the needle and the inoculation, we will proceed from the actual situation, and make self-made pipe and inoculation spare parts. These “soil spare parts†are better and “ The matching of foreign equipment has been successfully used in biotechnology fermentation production.
In short, the digestion, absorption and use of imported instruments and equipment in enterprises is an important task. No matter what problems arise in this process, you have to rely on your own efforts to find out more about the equipment, the working principle of the instrument and the design of the tube coils. The successful use of the US NBS40L fully automatic fermenter also illustrates this.
Experiences of using American NBS40L automatic fermenter
NBS40L fermenter is a large-scale automatic fermenter produced by New Brunswick Scientifi Company of the United States. It has the advantages of high degree of automation, convenient operation, small size and high degree of collection. The device has a stainless steel structure. The microcomputer controls the fermentation parameters and is well adapted to the fermentation production of biotechnology products.
Third, the problems and solutions in the sterilization process
(Chia Freda pharmaceutical Huhong Jie Wenzhong etc.)
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