International Journal of Research In Science & Engineering e-ISSN: 2394-8299

S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

Effect of Bubble Characteristics on Mass Transfer Coefficient in an Internal

Loop Airlift Bioreactor for Microbial Production of Daptomycin

Ipsita Chakravarty1, Sukhendra Singh2, Subir Kundu3*

1
Ipsita Chakravarty School of Biochemical Engineering, IIT (BHU), Varanasi
2
Sukhendra Singh School of Biochemical Engineering, IIT (BHU), Varanasi
3
Subir Kundu School of Biochemical Engineering, IIT (BHU), Varanasi (Corresponding Author)
1
ipsitachakravarty14@gmail.com
2
sukhendra86@gmail.com
3
skundu.bce@itbhu.ac.in

ABSTRACT
The present work focuses on the influence of the number of holes in a plate type sparger on the bubble size
distribution inside a 2L airlift bioreactor using image processing tools of MATLAB. Mathematical correlations
were used to predict the overall gas holdup, interfacial area and further the volumetric mass transfer coefficient
was estimated in the reactor for two different spargers based on the bubble size distribution. The effectiveness of
these correlations was validated by experimental data. The value of gas holdup of the multiple holes sparger was
higher than that in the single hole sparger. During fermentation, the KLa significantly increased from 30 h-1for
single hole sparger to 65 h-1 in case of multiple hole sparger. The bubble characteristics were effectively utilized
to study the hydrodynamics inside an airlift bioreactor and sparger geometry could be used to improve the oxygen
mass transfer characteristics of the fermentation broth and hence to ameliorate the production of Daptomycin.

Keywords: Daptomycin, Fermentation, Bubble size distribution, KLa, Gas hold up

-----------------------------------------------------------------------------------------------------------------------------

1. INTRODUCTION

Daptomycin is a modern-day anti-MRSA cyclic lipopeptide antibiotic produced by fermentation using

Streptomyces roseosporus, which is highly filamentous actinomycetes. The cell morphology of this producer
microorganism affects the hydrodynamics of the fermentation broth and hence, hampers the production of this
valuable drug. In our previous work with Daptomycin fermentation in Airlift Bioreactor, the shear thinning
properties of the fermentation broth was dealt efficiently through cell immobilization and morphological
modifications [1]. But, the issues of mass transfer inside the bioreactor are still unaddressed. Airlift bioreactors
have been widely applied in biochemical engineering due to their simple construction, low power consumption,
good mixing and relatively high mass transfer efficiency especially for the filamentous microorganisms and
their products[2]. Oxygen mass transfer plays an important role in the bioreactor performance especially for
aerobic fermentation processes. The volumetric mass transfer coefficient, K La, depends on the interfacial area of
gas bubbles and the physical properties of fluid. Bubble size and bubble number in the bioreactor have
substantial effect on the mass transfer [3]. Previously, researchers have investigated the local hydrodynamic
parameters of airlift reactors [4,5]. Sparger structure has considerable role in airlift reactors [6,7]. The influence
of reactor geometry and physical parameters have been studied in case of inverse internal loop airlift and
fibrous-bed bioreactor [8,9]. The effect of various hydrodynamic parameters with respect to different superficial
gas velocity and sparger structure for the design of internal loop airlift bioreactor is extensively studied in this
work. Empirical correlations are used to predict hydrodynamic parameters for the overall gas holdup and the
volumetric mass transfer coefficient in the reactor for different spargers. The efficacy of these correlations is
validated by experimental data.

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 International Journal of Research In Science & Engineering e-ISSN: 2394-8299
S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

2. MATERIALS AND METHODS

2.1 Bubble size and bubble number
A single hole plate type sparger and a multiple hole (N=5) plate type sparger of same diameter were taken. For each
of the spargers, the superficial gas velocity was varied from 0.001 Usg (m/s) to 0.005 Usg (m/s).

Figure 1: 1mm Single Hole and 1mm Multiple Hole Plate type Sparger
A photographic method was applied to measure the bubble size and bubble numbers using high speed DSLR
camera. MATLAB Image Processing Tools using MATLAB 8.6/R2015 b was used to process images at a scan rate
of 1000 frames per second to get the histogram for bubble diameter versus bubble numbers.

Figure 2: Scheme of Bubble Size Distribution Analysis

Between 1000 and 5000 bubbles were counted for determination of the size distribution, using 30 photographs. The
Sauter mean bubble diameter was calculated.

2.2 Gas holdup

The gas holdup was determined experimentally by the volume expansion method with respect to different
superficial gas velocity for two different spargers and also from the data obtained from bubble size
distribution.
ε= (1)

Where VG is the volume of gas bubbles in the bioreactor in m3 and VL is the volume of liquid media in the
bioreactor in m3. Volume of gas is actually the total volume of bubble multipled by number of bubbles of
a particular size. Volume of liquid is the volume of liquid media taken i.e. 1 litre.

2.3 Volumetric mass transfer coefficient

The interfacial area was calculated from the gas hold up values obtained from the bubble size distribution

a= (2)

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 International Journal of Research In Science & Engineering e-ISSN: 2394-8299
S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

d is the Sauter mean bubble diameter.

The trend consistency of interfacial area of the gas bubbles for each sparger was studied with the volumetric mass
transfer coefficient, KLa, measured by the dynamic gassing out method with respect to the superficial gas velocities.

3. RESULTS & DISCUSSION

3.1 Bubble Characterization

The bubble size distribution inside the airlift bioreactor is shown in Table 1. The results reveal that the sauter mean
bubble diameter is smaller for the multiple hole sparger than that for the single hole sparger under the same
superficial gas velocity.

Table 1: Bubble characteristics under different air velocities for different spargers.

Superficial Single Hole Sparger Multiple Hole

gas Sparger
velocity Sauter Bubble Sauter Bubble
Usg(m/s) Mean Number Mean Number
Bubble (1/L) Bubble (1/L)
Diameter Diameter
(m) (m)

0.001 0.0036 750 0.0025 2500

0.002 0.0038 1100 0.0028 3035
0.003 0.0039 1580 0.0030 3200
0.004 0.0042 1855 0.0036 4100
0.005 0.0044 2020 0.0038 4500

The bubble number significantly increases with increasing superficial gas velocity for both the spargers. The bubble
number of the multiple hole sparger is quite larger than that of the single hole sparger under the same superficial gas
velocity.

3.2 Gas holdup

The gas hold up values were calculated using bubble diameter and number of bubbles for each diameter as a
function of superficial gas velocity as depicted in Figure 3.The value of gas holdup of the multiple holes sparger was
higher than that in the single hole sparger. Small bubbles rose slowly and stayed longer in contact with the liquid
media which allowed enough time for oxygen to dissolve and distribute. Small bubbles produced in large numbers
led to higher gas holdup values. Due to higher number of bubbles generated in case of multiple hole sparger, the
overall gas holdup increases almost linearly with the superficial gas velocity.

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 International Journal of Research In Science & Engineering e-ISSN: 2394-8299
S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

Figure 3: Evolutions of gas hold up with superficial gas velocities for different spargers

At higher superficial gas velocity, bubble swarms and the bubble coalescence make flow pattern complicated,
resulting in slow rise of gas holdups. Similar results were obtained by Chisti [10] and Lu et al. [11]. As shown in
Figure 3, the overall gas holdup obtained with the multiple hole sparger is higher than that of single hole sparger.
Therefore, the larger number of holes lead to larger number of bubble formation and of small diameter which
attributes for an overall increase of gas holdup for the multiple hole sparger. Moreover, the smaller bubble size and
larger bubble numbers imply larger circulation of bubbles, leading to a higher gas holdup. The following equation
establishes the correlation between the gas holdup and the superficial gas velocity [10]:

ε=αUsgβ (3)

where α depends on liquid physical properties as well as the reactor geometry, and β is generally found to be a value
ranged from 0.4 to 1 [12].Fitting Eq. (3) to the experimental data of different spargers, the parameters determined by
the regression are listed in Table 2.

Table 2: Parameters of correlations between the gas hold up and the superficial gas velocity for different spargers.

Sparger type α β R2
Single Hole Sparger 0.0161 1.0274 0.987
Multiple Hole Sparger 0.0176 1.1478 0.996

These results were found to be similar to those reported by Chisti [1].The gas holdup estimated from bubble size and
bubble number was quite consistent with the experimental data [9,12] obtained using bubble characteristics.

3.3 Mass transfer

The specific interfacial area and gas hold up can be related as:
a= (2)

Putting the experimental values in the equation (2), a higher gas holdup and a smaller mean bubble diameter lead to
a larger specific interfacial area. Higher value of interfacial area is obtained for multiple hole sparger than single
hole sparger at a given superficial gas velocity as shown in Fig. 4. The specific interfacial area increases linearly
with increasing the superficial gas velocity. Since, K La is directly proportional to the specific interfacial area, the
increase of KLa arises mainly from the increase in the specific interfacial area. As the specific interfacial area can be
expected to depend indirectly on the gas holdup [13-15].

Figure 4: Interfacial areas with respect to different superficial gas velocities.

Experimental data of the volumetric oxygen transfer coefficient K La for different spargers as obtained from dynamic
gassing out method are plotted in Fig. 5. The data shows that the volumetric oxygen transfer coefficient increases
almost linearly with the superficial gas velocity for both the spargers.

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 International Journal of Research In Science & Engineering e-ISSN: 2394-8299
S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

Figure 5: Evolutions of volumetric mass transfer coefficient with superficial gas velocity for different spargers.

It indicates that the KLa obtained with the multiple hole sparger is higher than that of single hole sparger. During
fermentation, the KLa significantly increased from 30 h-1for single hole sparger to 65 h-1 in case of multiple hole
sparger [14]. As applied for overall gas holdup, the following equation is applied to correlate the volumetric mass
transfer coefficient with the superficial gas velocity:

KLa=αUsgβ (4)

The experimental parameters obtained for both spargers fitted well in the equation.

Table 3: Parameters of correlations between the volumetric oxygen mass transfer coefficient and the superficial gas
velocity for different spargers.

Sparger type α β R2
Single Hole Sparger 0.2021 0.5792 0.9945
Multiple Hole Sparger 0.2843 0.7426 0.9839

The superficial gas velocity directly affects gas holdup. The gas holdup further affects the specific interfacial area
and thus, volumetric oxygen mass transfer coefficient. However, the relationship between superficial gas velocity
and volumetric oxygen mass transfer coefficient is also influenced by bioreactor geometry, sparger designs, three-
phase mixing and flow patterns making it elaborate and complicated.

4. CONCLUSION
The influences of the sparger designs on hydrodynamics and mass transfer characteristics in a sparged internal loop
airlift bioreactor have been investigated for fermentation process. It is found that gas holdup and volumetric oxygen
mass transfer coefficient is strongly related to the sparger design and especially the superficial gas velocity. The
comparison between experimental data of the single hole sparger and the multiple hole sparger indicated that the
sparger with the larger number of orifices was more efficient for the mass transfer, because it produced a smaller
mean bubble diameter and larger number of bubbles leading to greater gas hold up values as compared to single hole
sparger. Moreover, for a given sparger structure, the effect of the superficial gas velocity on the K La was mainly
reflected on the specific interfacial area as calculated using gas hold up values. Empirical correlations could predict
the hydrodynamic parameters for the overall gas holdup and the volumetric oxygen transfer coefficient for different
spargers. The empirical correlations had satisfactory agreement with the experimental results obtained. Therefore, it
could be concluded that the bubble characteristics could be utilized to study the hydrodynamics inside an airlift

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 International Journal of Research In Science & Engineering e-ISSN: 2394-8299
S CHEMCON Special Issue : March 2018 p-ISSN: 2394-8280

bioreactor and sparger geometry could be used to improve the oxygen mass transfer characteristics of the
fermentation broth and hence to ameliorate the production of Daptomycin.

ACKNOWLEDGEMENT
The authors acknowledge the financial supports from MHRD and School of Biochemical Engineering,IIT ( BHU),
Varanasi, India for providing laboratory and technical support.

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